Podcasts about cpy

ATMakers emailed and asked if we'd make a Trinkey (https://www.adafruit.com/search?q=trinkey) with a TRRS jack (https://www.adafruit.com/product/5764) on the end, for making customizable HID devices that use simple 3.5mm audio jack connectors. ideally something that will make AT - Accessibility Technology - projects EZ to build and maintain. here's our take on it, what's interesting is each pin of the jack - including the two switches for tip and ring1 - connect to GPIO / analog inputs on the SAMD21. there's no pre-defined VCC or GND because different switch and joystick makers have different configurations and we might want to change around what pins are power, ground, pullups, etc! this board will be great for use with CircuitPython to make any sort of AT switch or potentiometer interface super fast, to demo it we've written up a quick CPy sketch that will listen for two switches, and when they're pressed, blink the NeoPixel and emit keypresses as if its a keyboard. code is in the PR for this board (https://github.com/adafruit/circuitpython/pull/9190#issuecomment-2067764251) since we have great HID support it could also be a mouse or joystick device and it will work with any computer/tablet/console with a USB port. Visit the Adafruit shop online - http://www.adafruit.com ----------------------------------------- LIVE CHAT IS HERE! http://adafru.it/discord Subscribe to Adafruit on YouTube: http://adafru.it/subscribe New tutorials on the Adafruit Learning System: http://learn.adafruit.com/ ----------------------------------------- #adafruit #atmakers #accessibility

Join Skit, Adam and Scot for this week's lively and energetic episode of YAMology where Alex and Matt call eachother by the wrong name for half the pod. The trio (we) get into the IST and Awards for MVP, 6MOY, ROY, CPOY, DPOY and CPY, obviously pronounced correctly in YAM fashion. We also have an overtime section where we touch on the WNBA and NBA draft prospects. Enjoy and sending Hugs and kisses to all you Northern Hemisphere folk from YAMology Pod. (Please note that this description was written by Matt, the other co-hosts may or may not share the same opinions.)Timestamps0:00-5:40 Intro + Scouting NCAA Basketball in person5:40 In-Season Tournament Takeaways16:35 NBA Awards Discussion Clutch Player of the Year (CPOY)21:04 Rookie of the Year (ROY)27:01 Sixth Man of the Year (6MOY)32:35 Most Improved Player (MIP)39:15 Defensive Player of the Year (DPOY)42:15 Coach of the Year (COY)46:15 Most Valuable Player (MVP)55:06 2024 NBA Draft Prospects Linktreehttps://linktr.ee/y4m0l0gyhttps://www.youtube.com/@YAMologyNBATwitterhttps://twitter.com/Y4M0L0GY

Making your mind a safe space can be hard, especially during a busy audition season. This week, Alyssa sat down with licensed therapist Madeleine Dopico to chat about how to be kind to your mind during stressful times of your life.  Don't forget to check out friend of the pod Ashlee Waldbauer and her work on Nevertheless Schlee Persisted wherever you listen to podcasts! Madeleine's Information: Website LinkTree  Instagram @iamdopico Spotify Apple Music YouTube  Madeleine Dopico, LMSW is a mental health therapist + musician/performer in NYC. Before her clinical training, she was the CEO of a mental health tech startup and recently launched “The Mental Health Party” - an organization aimed at fostering mental health positivity and community. Madeleine is striving to weave mental and emotional wellness more seamlessly into our days, through efforts like the “Mental Health Integration” collaboration with Core Power Yoga UWS. A passionate CPY Yogi herself, she is exhilarated to be working with CPY teachers to bring a cohesive mind and body experience to all those interested in personal growth and exploration! Resources: Budget Bundle 2.0 Learn Your Music The Right Way Workbook Goal Slaying Planner Let's get social: Support the podcast Follow us on social media Search our website Check out our Shop Follow Ellen Follow Alyssa --- Send in a voice message: https://podcasters.spotify.com/pod/show/modern-singer-llc/message Support this podcast: https://podcasters.spotify.com/pod/show/modern-singer-llc/support

The lads are back after a 12 day break - squeezing in the episode the day before the deadline like an Evan Ferguson goal at the end of a game (and then getting injured....).- We discuss all the new players and how we wanted to pick Mudryk, but cant because hes a terrible person.- We are still United IN and Andy bets Dom that United will win vs. CPY and LEE- We are full on Everton targeting, but think Dyche will park the bus and pray

This week, Workout Woman aka Ashley Fern (@disco_infern0) shares her latest workout review. The episode kicks off with Aleen and Ashley catching up and then they share their DSTW of the week. Ashley's is about finally hitting a workout goal she was striving for and Aleen's is about getting rid of things from her closet that were no longer serving her. Aleen then reads a listener DSTW about learning to be more comfortable with wearing crop tops and how the listener was able to overcome her internal dilemma in time for a big performance. They then get into the workout report of the month, which came highly recommended: CorePower Yoga. To start off, they address the bad press that the company has gotten, but remind DST'ers that this review is about the workout and not the business itself. Ashley then breaks down what her class experiences have been like at different CPY studios, if it's worth going back, and her overall thoughts on the workout aka if the workout is worth needing to wash your hair. Don't forget to submit your recommendations for workout classes for Ashley to try by dming @dietstartstomorrow or emailing dst@betches.com.

Join Scott for his final Deep Dive before paternity leave. During the dive he answers questions, shows a PyUSB example and chats with @foamyguy who will be streaming in the Deep Dive slot while Scott is out. Check out FoamyGuy's other streams: https://www.twitch.tv/foamyguy_twitch https://www.youtube.com/channel/UCDfd9ZAcB8kpa3pyEQwLXVA Next week will be Deep Dive with Foamyguy at 2pm Pacific / 4 pm Central. Support Scott and foamyguy by buying Adafruit hardware. - http://www.adafruit.com All notes are available on GitHub with links into the videos. Thanks to @askpatrickw and @dcd for making and maintaining the notes. https://github.com/adafruit/deep-dive-notes/ 0:00 Getting started and hellos 1:30 is this the last one? 4:51 it's 2pm - let's get the show on the road 6:00 Housekeeping 7:10 Adafru.it/discord 8:15 foamyguy next week 11:00 github for notes https://github.com/adafruit/deep-dive-notes/ 13:15 Any tips for an electrical engineer that wants to be a hardware development engineer? 14:46 Are We building a whole usb phy stack in Python? 16:08 Last week - pyusb api in Circuit python 16:35 Tinyusb 17:25 learn.adafruit.com great resource - 20:05 good progress with usb host 22:07 usb/basic_mouse.py 24:33 pyusb basic example import usb.core 25:37 embedded C / microcontroller history / flight controller / dataflight / cleanflight drone / IMU 27:30 flight controller overview / baseflight / forked into cleanflight / forked into betaflight 29:45 Chickadee tech http://chickadee.tech/ 31:49 Polystack 32:32 transition to adafruit 33:28 “It was really interesting following his struggles on Show&Tell!” 34:37 any experience with telecom or signals ? what do you think about that kind of stuffs? 35:22 any CPy quadcopters out there? 36:56 “what's my hobby now that my hobby became my job” 38:00 https://adafru.it/discord 40:00 FPGA ideas for CP 41:00 do we have an FPGA channel on discord ? #fpga in discord 43:44 back to usb.core USB_VID and USB_PID 45:10 using CP to patch the FPGA 46:30 convert ID to strings and disconnect from kernel 47:15 loop on endpoint 0x81 47:38 lsusb 48:21 What is a boot mouse? / Boot Interface Subclass 49:47 endpoint 0x81 in 50:45 https://wiki.osdev.org/USB_Human_Interface_Devices 52:31 intellikey overlay - translating key presses using CP 54:15 starting with assuming that CP knows a lot about the device 55:50 teensy 4 and imx 1060 and rp2040 56:30 run the script 57:33 connect with foamyguy 59:10 foamyguy streaming over the past year 1:00:48 circle due to OBS and other meetings - virtualcam into teams 1:10:50 PopOS- but started on Windows ( https://pop.system76.com/ ) 1:02:58 foamyguy programming background 1:08:55 original rPi had instructions to accelerate java 1:12:55 RP2040 1:13:33 don't move away from google fiber :-) 1:14:35 seems like Damien really changed the game with Micropython? 1:15:20 BBC microbit was first experience with embedded python 1:17:25 Edge badge was first Hackaday CP device with CP - 1:21:00 Secrets.py transition to secrets.json, but toml has comments! 1:22:15 environment variables for CP 1:22:45 ​jsonc allows comments 1:25:16 interactive tablet application - more flushed out than py-portal 1:30:00 https://play.date/ 1:32:45 https://www.steamdeck.com/ 1:33:00 zync fpga zybo 1:38:21 blinka and ML / tensorflow lite 1:40:40 Digital Signage microprocessors - hard to compete with raspberry pi 1:43:53 https://www.screenly.io/ 1:46:12 foamyguy youtube stream content - similar to deep dive 1:48:00 Are you still going to do the saturday stream (10AM Central time) or are you moving this to the Friday deep dive. 1:49:18 interest in Async io as related to display io 1:50:48 CP Parsec noted 1:51:50 foamyguy working for Adafruit on mondays 1:55:53 android - ble workflow - glider app on ios 1:58:35 python editing experience / type enabled editing 2:02:27 https://www.twitch.tv/foamyguy_twitch 2:02:49 https://www.youtube.com/channel/UCDfd9ZAcB8kpa3pyEQwLXVA 2:04:32 wrap up adafru.it/discord 2:07:02 cat cam / pet the cat 2:08:00 end fo stream ----------------------------------------- LIVE CHAT IS HERE! http://adafru.it/discord Adafruit on Instagram: https://www.instagram.com/adafruit Subscribe to Adafruit on YouTube: http://adafru.it/subscribe New tutorials on the Adafruit Learning System: http://learn.adafruit.com/ -----------------------------------------

Join Scott as he recaps ESP BLE work, keyboards and USB host. Support him and Adafruit by purchasing Adafruit gear from https://adafruit.com. Join the Discord at https://adafru.it/discord 0:00 Getting Started 5:38 Housekeeping 10:00 Mailbag 10:55 installed the CP 7.1.1 on a Raspberry Pi Z W2 issues. / ( use latest ) 15:03 - overhead / return to mailbag 15:42 AF Data cable /4449 https://www.adafruit.com/product/4449 17:20 Multifunctional USB Safety Tester https://www.adafruit.com/product/4232 17:50 Usb CP2102N usb to serial converter / 5335 19:10 New products from UM ESP 32 Feather S3 https://esp32s3.com/ 22:20 Tiny S3 24:23 Pro S3 / PicoBlade to JST-PH 28:59 BLE - converting broadcast net ( NRF52 read sensor and broadcast ) w/bridge 31:30 Desktop / gthub CP BLE_BroadcastNet 32:05 Can an iphone work as ble key for our iot devices? key like authentication key 33:03 PR for Add native bridge example 34:01 io.Adafruit 34:22 Raspberry PI and the stock shortage that seems to have created a 50+ dollars Pi zero a thing. 35:05 BLE bridge with AIOBLE? Or similar? All Asyncio? 35:31 Bridge core ( ble.start_scan ) bls_broadcstnet*.py 37:37 adafruit io bridge dashboard 40:28 Adafruit LED plastic enables visibility of the purple blink 42:20 Bridge output 42:32 so the background BLE scans are non blocking so CPy user code is not interrupted? 43:35 Blinkng 2 different colors to know the different sensor data received 45:03 Color picking trick rainbow.colorwheel based on reversed_address 47:55 Native bridge is not in the library yet - see Learn Bluetooth LE sensor Nodes to Raspberry Pi WiFi Bridge (by John Park ) 49:45 complicated instructions to get the WiFi Bridge going 50:32 Packet Buffer examples working now PR 158 - insulate code from time sensitivities 52:15 Do you have WiFi always on, and connected to the router? Do you know your current (I) use when you are scanning and sending with WiFi? 53:50 ble_packet_buffer_client.py 55:45 ble.start_scan 56:21 NRF app for BLE advertising 58:40 how do you make a custom BLE server *buffer_service.py complex characteristic ( perhaps https://circuitpython.readthedocs.io/projects/ble/en/latest/characteristics.html ) 1:04:35 Adafruit BLE characteristics ( ble io bindings ) 1:07:38 download the two files from S3 the uf2 and bin, I need to write the bin first to the esp32-s3 then it should boot with a bootloader directory where I drop the uf2 file in. Is that correct? 1:09:35 data descriptors in python https://docs.python.org/3/howto/descriptor.html 1:11:00 Is there a way to make a raspberry pi zero run circuitpython the same way the pico does? I am trying to use usb_hid on the zero but it wont work and pico runs to slow 1:11:39 learn guide bare_metal no os https://learn.adafruit.com/circuitpython-on-raspberry-pi-bare-metal-no-os 1:13:17 USB Host Pico-PIO-USB using PIO https://github.com/sekigon-gonnoc/Pico-PIO-USB 1:19:00 Pico usb host demo 1:21:57 data + and - wires go to uart tx rx pins or any pins ? 1:24:41 github issue 5986 1:26:44 Seniply / colemake configurator qmk.fm 1:27:19 switch to overhead 1:27:45 ​In order to have a Pico or ESP32 communicated with a user space program, is Serial COM the only option? 1:28:29 ​So I followed the guide and have CircuitPython installed (no os) on the raspberry pi zero 2. But it doesn't seem to work. But my pc doesn't detect anything when I plug it in 1:29:14 interested in making a midi usb host. I hope it's not so different than what we are doing here 1:30:00 ETA of usb 2.0 HOST support for HIGHSPEED devices in TinyUSB for original rpi zero (BCM2835) ? (only Teensy 4 chip has both supported for now) 1:31:33 flashing rainbow on hdmi what does that mean? 1:32:27 TinyUSB has USB Host support for HID (keyboard,mouse) and there's a PR for MIDI, but that's it for now I think 1:32:48 pluki (sp) trackball interaction with keyboard 1:34:00 layers on CMK Configurator 1:35:10 running custom code ( not CMK or QMK ) - for colmax keyboard 1:36:02 how long did it take you to make the switch? 1:36:25 if you want more ergonomic keyboard - don't use QWERTY 1:37:55 Split keyboards are the way to go 1:38:45 keyboardio keyboard ( previous keyboard ) 1:42:58 pimpmykeyboard.com 1:44:50 Keycap families DCS, DSA, and SA Family - ortholinear 1:46:35 keyboard can be CP or HID only 1:47:00 code.py in the keyboard 1:49:15 future refactoring… 1:49:40 “this is a bug” - editing the keyboard code using the keyboard - risky 1:52:25 other standards via and vial 1:52:32 is adafruit going to make a diy vr controller as they HAVE made a vr headset and i have it but im using terrible controllers and i was wondering if you would make some. 1:54:33 keyboard “stages” 1:57:45 any ideas for naming ? 1:58:05 Broadband update / https://wa-law.org / right to repair / share the cities slack 2:05:47 wrapping up / next week also friday / USB Host / https://github.com/adafruit/deep-dive-notes 2:08:30 pet the cat 2:09:14 have a great weekend

Join Scott as he recaps other folks' #CircuitPython2022, discusses ongoing ESP32-S3 work and answers questions folks have. Support Adafruit by purchasing hardware from https://adafruit.com Chat on the Adafruit Discord at https://adafru.it/discord. All notes are available on GitHub with links into the videos. Thanks to @askpatrickw and @dcd for making and maintaining the notes: https://github.com/adafruit/deep-dive-notes/ 0:00 Getting Started 1:00 hellos to all 3:00 cat cam 3:40 Hello/housekeeping 7:00 github notes https://github.com/adafruit/deep-dive-notes/ archive 9:09 CP 2022 - send emails - get blogged 10:07 Kevin - display io 11:25 Q: when debugging with print statements, is there a way to include the statement line number in the print string? 11:45 Question: does circuit python have the ability yet to play simultaneous audio? E.g. via 2 wav files or foreground / background. Maybe through mixer 12:58 Desktop - github - why is issue 5881 is "longterm"? https://github.com/adafruit/circuitpython/issues/5881 (BLE ) 14:45 github BLE Adafruit_CircuitPython_BLE 17:25 desired: want to disconnect the undesired centrals! / examine the source code 19:30 Various issues involved - connection central vs. peripheral 20:55 github Adafruit_CircuitPython_BLE issues / look into device info service 22:30 back to the 2 wave files 23:53 Q:working with the analog in features on a QT pi 2040 and when I tied the pin I was reading from to ground on the board it was giving me a value other than zero, around 160. Is this normal? 24:45 Q: built latest CPY for S3 - doesn't boot on any of my S3 boards. Known issue? copy firmware.uf2 over, seems ok, re-starts, never mounts. No error on UART0 output. 25:48 back to CP 2022 - graphical widgets 28:22 unpythonic.net 31:05 github gist / broadcom / midi 32:09 forums / 33:00 blog comment - 33:34 BlueMicro / skip forward to CircuitPython 2022 / seesaw / i2c peripherals / keyboards / very deep sleep 36:00 - RISC-V vs. ARM Core… 36:45 Awesome CircuitPython called Popular Projects Enabled by CircuitPython 37:44 forums / overclocking / platform knowledge 39:00 SPI peripheral 39:25 forums / CP.org login / my adafruit page ? 40:00 intuitive flash file system / handle secrets storage 40:53 limited multicore access 41:20 Q: ​if any of your viewers could please, please, take up i2c peripheral support for the rp2040 I would be grateful! 41:57 I2C peripheral thoughts - copy API 42:55 integrate learn guides with what user purchased 44:00 Unified event system 44:33 some comments on youtube re: CP 2022 45:20 github gist 46:50 Lego collaboration? 47:35 Python threading 48:20 considerations when setting priority 48:53 adafruit blog 50:30 Multicore 52:07 ulab extension 54:30 new CP podcast coming 55:20 I played with the Micropython multicore a few months ago and it wasn't stable 55:39 dafruit_requests and adafruit_minimqtt would be nice to be threaded / asyncio 56:15 AIOHTTP Async IO for networking 58:30 Does Wippersnapper use mqtt? 59:10 be more sciency 1:00:20 2022 will be different with weeks off from Deep Dives 1:01:49 switching gears to ESP-land 1:02:00 Adding BLE to ESP 1:03:30 How many cores are on the ESP32-S3 ? 1:05:11 tannewt esp_ble branch 1:06:49 beyond compare / sublime merge ( git manipulation tools ) 1:07:35 How much power does the ESP32-S3 use with BLE (no WIFI)? 1:10:10 Are there any examples in CP doing double buffering aka glitch free bucket logging ? Maybe via M2M ? 1:10:35 how well does the nordic power tool work with measuring other processors power usage? 1:12:09 also the JouleScope 1:12:55 any information if there are nRF52840 successors on the horizon with more RAM memory? 1:13:17 NRF 5340 1:15:00 Zepher support not in Circuit Python 1:15:18 rf 9160 1:15:39 dual M-33 1:16:30 https://www.youtube.com/watch?v=JFDiqPHw3Vc&ab_channel=AndreasSpiess 1:16:49 The second core is basically just a built in HCI adapter so if you can get the IPC to work on bare metal, you don't need Zephyr. 1:17:50 Bluedroid and Nimble ( grew up alongside Zepher ) 1:18:25 github nrf5340 apache mynewt nimble 1:20:11 back to esp_ble branch 1:23:20 start with scanning, advertising .. 1:25:08 talk over core issues 1:25:40 Pull requests 1:26:19 pr 5902 1:30:12 pr 5899 1:31:06 pr 5860 1:31:25 pr 5884 1:32:32 pr 5885 - first CP multicore bug fixed by tannewt 1:36:30 pr 3892 ESP NeoPIxel fixes / resetting pins 1:40:20 lower power when pins are ‘pulled' vs. floating 1:42:20 looking for the low power pin state pdf https://www.st.com/resource/en/application_note/an4899-stm32-microcontroller-gpio-configuration-for-hardware-settings-and-lowpower-consumption-stmicroelectronics.pdf 1:45:06 pr 5894 1:45:50 C3_writeable ports/esprif make BOARD=espressif_ep32s3_devkitc_1_n8r2 1:51:30 ESP32-S2-DEVKITC-1 on digikey 2:04:53 wrap up - follow on discord https://github.com/adafruit/deep-dive-notes 2:06:59 pet the cat 2:07:03 Have a great weekend ( timecodes may have drifted by 30 seconds or so )

I'm sponsored by Adafruit to work on CircuitPython. Support them, and by extension me, by purchasing hardware from https://adafruit.com Chat with me and lot of others on the Adafruit Discord at https://adafru.it/discord. Deep Dive happens every week. Normally Fridays at 2pm Pacific but occasionally shifted to Thursday at 2pm. Typically goes for two hours or more. Questions are welcome. Next week will be on Friday as well. 0:00 Getting started and hellos 0:04:44 It is 2 01 - welcome and Housekeeping 0:08:27 Circuit Python 2022 introduction - long term direction 0:09:30 documented purchasable kits for kids/teens (see learn guides, adaboxes, shopping gift guides) 0:13:21 What is the underlying OS in Circuit Python ( ESP IDF build on Free RTOS) 0:14:40 Does CP ( Circuit Python ) on Arduino 0:15:08 Is the Raspberry Pi 2W SD card image working if you flash it from scratch? 0:16:10 Pull requests #5800 (initial Pi Zero support) https://github.com/adafruit/circuitpython/pull/5800 0:17:36 ESP-32 (not supported) see ESP-32 S2 and S3 0:18:15 conferences ? ( deep dive notes ) 0:20:40 search available for deep dive notes ) 0:21:35 CP advantages ( over Uno ) 0:23:02 join discord here ​https://adafru.it/discord 0:23:20 CP repl - compared to auto reload 0:24:31 wrap up recap - ( add Piunora and Zero ) 0:25:23 Open issues 0:26:12 Broadcom support corrupts SDCard #5706 https://github.com/adafruit/circuitpython/pull/5706 0:27:35 SD Card emulator? 0:29:07 Neopixel support on PWM capable pins #5727 https://github.com/adafruit/circuitpython/pull/5727 0:29:55 Upcoming - S3 support 0:30:25 4x64 Led Matrix using Matrix_Portal_Animated_Gif 0:33:00 no keyboard on the Pi400 because it require to be USB host? 0:33:25 Pi vs Pi Accessories 0:35:22 Wifi on the Pi - MicroPython support 0:35:35 S3 interest in BLE workflow 0:36:40 What do I do with this old Pi 0:37:13 What do you prefer in terms of IDE or text editor? 0:39:30 Where do I start if I want to get involved in contributing to CP ? ​https://circuitpython.org/contributing 0:41:14 Legaslative non-work topic(s) right to repair, broadband 0:42:12 Circuit Python 2021 0:42:35 Did you get into the QMK config tool output thing? ( not yet) 0:43:44 twitch - best board to get started with CP 0:45:50 Pico at the top of the Downloads page since the beginning! 0:46:10 Low power 0:46:55 BLE WiFi provisioning 0:47:20 Should the old ESP32 regain support now that there is a BLE workflow? (hope so) 0:48:45 is it possible to run (execute) another .py file before code.py? ( boot.py runs before USB is active ) 0:51:18 Circuitpython 2022 - look at 2021 0:51:55 is that HDMI? how do you change the resolution on RPi? 0:58:00 tiny logic analyzer project… 1:05:30 how to run something besides code.py 1:12:13 Are there any plans to support different pictures/graphics types (PNG, JPG, SVG) other than BMP? in CP? 1:15:50 Could auto parse settings.toml at boot and pass as environment variable 1:17:15 TOML toms markup language 1:26:40 TOML has comments 1:27:00 Would there be a way to have one RP2040 firmware and have board variations that do not require their own firmware? Like a config file or something small that just include the variation between board? 1:33:00 Bryans rules for good documentation 1:34:41 CP 2022 kickoff post / 2021 at https://blog.adafruit.com/2021/01/15/tannewts-focus-for-circuitpython2021/ 1:42:15 - the chip shortage causes CP to be flexible 1:44:02 Round LED screen ? 1:48:56 Love to hear your thoughts on CP 2022 - email circuitpython2022@adafruit.com Or https://blog.adafruit.com/2022/01/01/circuitpython-in-2022-circuitpython2022-circuitpython/ 1:52:35 https://wa-law.org/ 1:54:30 slack for repair.org https://www.repair.org/ 1:55:05 ​I'd like to see the option to choose "CPY" with and without OTA partitioning... I get asked all the time why my boards that have 4MB of flash only have 900k available in the FS in CPY. 1:59:44 calling it a day - next week CP 2022 2:00:53 MP (will be very soon) uf2 friendly. so just drag MP onto UF2 like CPY! No more stomping CPY with MPY ! 2:01:30 Cat cam 2:02:02 Visit the Adafruit shop online - http://www.adafruit.com ----------------------------------------- LIVE CHAT IS HERE! http://adafru.it/discord Adafruit on Instagram: https://www.instagram.com/adafruit Subscribe to Adafruit on YouTube: http://adafru.it/subscribe New tutorials on the Adafruit Learning System: http://learn.adafruit.com/ -----------------------------------------

Scott recaps work on the Raspberry Pi running CircuitPython and then continues working on SD card support. Visit the Adafruit shop online - http://www.adafruit.com Chat with me and a lot of others on the Adafruit Discord at https://adafru.it/discord. Deep Dive happens every week. Next week is on Friday at 2pm Pacific. 0:00 Getting Started 09:32 Housekeeping 13:00 Raspberry Pi Zero 2 W discussion 19:50 raspberry pi .org vs. .com 20:51 Dune and DOS 21:49 USB Host and Tiny USB 23:07 Edit code on a Pi over BLE 23:38 working on Pi 400 steps to figure out 24:14 Pi-DOS project mention 26:00 Glasses come off / glasses setup 27:00 Pi Zero - epic for CP with .5 GB RAM 29:00 Zero 2 is the ultimate board for CP!!! 29:50 Review last week's status - 30:00 Now Tac has High Speed working 31:08 Last week - HDMI was very slow - investigate caches off/on 31:38 Not caching “enough” 32:48 Scrolling demo, much faster than last week, caching on for everything 33:42 Tweek resolution 34:00 “what is CP” 35:15 Still Broadcom chip 36:14 Flash speed vs. run from RAM 37:37 once I get the SD card working, there will be some re-organization for all the boards 38:20 experiment with screen resolutions, to demo refresh performance 40:17 640x480 REPL - pretty quick 42:00 1080p seems blurry 43:10 Are there any plans to make this a simpler process, eg moving it to a Java IDE? Suggested https://codewith.mu/ 45:10 check out visual studion circuitpython plugin 47:11 Python 3.10 has deprecated “distutils”. Spent my day on that for Yocto Project. It's nice to not need to worry with CP 48:00 rebooted to ‘lower' resolution 48:30 flash wear discussion Nand flash, Nor flash, flash protocol 49:36 Should you have some knowledge of Python before learning Circuit Python? 50:00 Pi4 is ARM64....so theoretically you could port CPY to the M1 chip 50:32 core of CP is just like Python 50:57 SD Cards technical proprietary, but … 51:27 Why use bare-metal CP over blinka ( ease of setup, just does one thing ) 52:20 Implementation 53:11 adafruit_sdcard.py - SPI 1, 4 or 8 bits at a time 54:15 here is a lot of IO stuff that you couldn't do under a Linux kernel like bitbanging is quite hard/limited in a non realtime OS 55:16 raspi3-tutorial / 0B_readsector / sd.c 56:39 habits that lead to more reusable code :-) 57:04 declaring 5 global variables on one line with no comments 57:37 SBD generated structs vs. 58:05 multiple things on one line - suggest using curly braces freely 59:00 initializing some but not all variables ( on one line ) 59:26 ‘weird' globals 59:39 q:is there a native async library in circuitpython? I found a library called "asynccp" it works for me now but it would be better to use the native way. 1:01:04 use single letter variables sparingly 1:02:06 perhaps run it through a formatter 1:02:18 “I still don't know what this code does” :-) 1:02:55 consider naming style for global variables 1:03:15 ‘circle' reference gighub rsta2/circle - well commented 1:04:10 check out the license 1:06:04 sdcard.org PDFs 1:06:44 Scott's Pi Zero arrives on Monday! 1:08:11 640x480 HDMI raspberry pi bare metal REPL demo - last piece is SD card reading in CP - connect to USB mass storage 1:09:40 goal: read SD card over USB 1:10:29 Exception levels - switch from EL2 to EL1 ( os exception level ) CP is running EL1 1:12:00 Waiting for high speed to be merged into tiny USB 1:12:40 HDMI output used for display IO 1:14:40 Looking at board.c in CP ports/broadcom/boards/PI4 1:15:15 sdioio API 1:15:35 “with this big chonky font, CP needs to implement CBM ASCII to get cool map-building "letters"” 1:15:42 detour - fantasy console - mimic but modernize 1:16:34 nerd fonts project (nerdfonts.com) 1:18:00 Does circuitpython use Unicode strings? 1:18:14 Twitter emojis opensourced twitter/twemoji 1:19:44 displayio doesn't display it yet - though emoji variable names do 1:24:30 in cpython you can only use unicode characters in variable names if the belong to the "letter" class, so you can do accented characters, or Chinese, or Hebrew, but not emoji 1:25:37 back to sdioio/SDCard.c 1:27:54 second argument to SD send command - refer to adafruit_sdcard.py 1:30:50 SD Specs - commands , and back to adafruit_sdcard.py 1:39:03 bztsrc/raspi3-tutorial 0B_readsector/sd.c sd_cmd() / 32-bit commands vs BCM2835 ARM Peripherals.docx 1:41:21 Q: been experimenting with the sdcard module on circuitpython but it stops the code if there is no card in the reader 1:42:36 consider the response type ( number of bits ) 1:43:54 sdioio/SDCard.h 1:45:03 sdcard.org Part1_Physical_Layer_Simplified_Specification_Ver800.pdf r2 response codes 1:48:57 autogenerated SVD generated file bcm2711_ipa.h 1:52:45 sdcard.org Design Guide « Whitepaper » is a pretty cool resource. https://www.sdcard.org/cms/wp-content/uploads/2020/11/SD_Express_Design_Guide.pdf 2:04:00 CM4 Appendix B - ordering codes ( wireless, eMMC, RAM ) 2:06:15 Wrap up - next week Pi Zero on Friday 2:09:35 have a great weekend

Here's another episode of Conversation with Friends with a different direction. You can watch the video version of the same episode at https://youtu.be/UYqDm-9-vAU As part of the celebration for AAPI Heritage Month, we have Calvin & Theresa Curameng from College Park Yoga, located in College Park, FL, a community just outside of Downtown Orlando. 20 years ago, it was not an easy decision for an Asian American person to embrace the life of an Urban Yogi. We went into business, medicine, technology, law or education. You get the idea. So when Calvin & Theresa Curameng whom the students affectionately refer to as C & T set out to create College Park Yoga, it was quite an adventure! The studio was built on "pizza boxes". (You will have to watch the interview to find out what that really means!) In 2020/2021, College Park Yoga encountered COVID-19. Like many other small businesses, CPY faced and is still facing an uncertain fate. Perhaps this is what yoga practice does for you: flexibility. This has also been a time full of newness and creativity for them. Calvin & Theresa created the Liquid Fire Yoga Practice: "Yoga practice is the art of learning to handle friction. The friction of the water and fire inside of our very being. The friction of opposition; action and inaction. It's the power to discern what you love from what you think you love." This is a quote from their website www.LiquidFireYoga.com You can find out more about who they are, their classes (online and in person) and many other fun and wonderful things they do by visiting their website: www.CollegeParkYoga.com #AAPI #AAPIHeritage #Yoga --- Send in a voice message: https://podcasters.spotify.com/pod/show/shirley-wang6/message Support this podcast: https://podcasters.spotify.com/pod/show/shirley-wang6/support

Today I'll answer questions and discuss the work to get a BLE workflow going in CircuitPython. I’m sponsored by Adafruit to work on CircuitPython. Support them, and by extension me, by purchasing hardware from https://adafruit.com Chat with me and lot of others on the Adafruit Discord at https://adafru.it/discord. Deep Dive happens every week. Normally Fridays at 2pm Pacific but occasionally shifted to Thursday at 2pm. Typically goes for two hours or more. Questions are welcome. Next week will be on Friday and will be the 1 year anniversary! 0:00 Getting started after technical difficulties 4:04 hello and thanks to 009 for the gpu clue 7:04 q: just started my first BLE project that i showed on S&T. i'm confused what the BLE components i need to setup some kind of "clear/reset connections and repair from scratch" button to make my board useable across devices without having to manually disconnect before reconnecting 9:19 core modules ble erase_bonding… 9:59 working on a digital thermometer for an old bmw that would obviously show the engine's temp. I want to use an stm32. 15:04 BLE - Bluetooth Low Energy 17:51 Circuit Playground Bluefruit - Bluetooth Low Energy 20:14 what applications would you recommend BLE for? 25:49 Bonding 29:45 I know you can SEND data with the Micro:bit BLE, but can you program it from mobile through BLE?? 30:14 Serial/UART over BLE is not "super standard" as opposed to Serial on Bluetooth. There is a circuit python LIB that does the Nordic way. But you cannot change the UUID for another copy of serial. 30:49 BLE workflow - discovery 35:59 for devices that allow multiple devices to connect (like a mouse that toggles between machines) do you need a separate BLE chip for each device? 37:04 Is this workflow tied to BLE or is it standard for regular (older) bluetooth as well? 37:24 The number of pairs, bonds, or connected? 40:19 ​I want to make a BLE keyboard that I can quickly switch between my phone, desktop, laptop, and RPi. So, what's the most bonds possible? 42:14 what security things should be we aware of when working w/ BLE? 46:24 Desktop / “where scott is at”... 46:54 Packet buffer fixes #4535 48:34 ​Don't the packets have sequence numbers? 49:21 Fix central pairing #4522 51:45 Proof of concept code for CP future behavior 52:28 I noticed a 7.0.0 tag has appeared in the CPY repo. wonder what that will bring 54:09 So it is not going to be in the "kernel" but in "library"? Or it is a mockup in "userland"? 54:39 Adafruit_CircuitPython_BLE_Creation / Creation IDs on github 58:04 instance id? ( NO ) 1:02:48 Creator IDs, how are you allocating them? So 0x1234_xxxx could be registered by one person/organisation? 1:03:56 Examples ( advertisement / scan? ) 1:05:04 File Transfer 1:06:54 The BLE SIG is in Kirkland, WA if you want to chat with them. I might have some contacts. 1:07:09 There is/was that file transfer solution over the REPL for micropython. Can you reuse that? 1:07:55 Do you envision the BLE file transfer protocol requiring the storage to be mounted writable from boot.py on the CircuitPython device? 1:09:54 ampy == That was the recommendation for micropython… ( “No - don’t like it”) 1:10:14 This might be old information... but at one point iOS didn't support serial over bluetooth. (Ham radio control apps used this). Has that changed ? 1:11:04 What is BLE range on CPB/Micro:bit (estimate) 1:12:29 Does BLE keyboard use scan codes or ASCII? ( Same as USB ) 1:13:46 BLE file transfer library 1:14:34 BLE number set 1:17:34 ​I'm going to need a way for my keyboard to know whether the device it's talking to is in Dvorak or US mode. 1:20:09 Flow Control 1:21:49 demo 1:23:09 In "Bluefruit Connect" you have packet based protocol, over serial with function like picking a color or sending image... you could expand that. 1:24:53 Is the bug that it's starting to corrupt the file system, but not finishing the job? Or that it's corrupting the filesystem at all? 1:25:19 Maybe that "Bluefruit Connect" and protocol and serial was there for old way to connect serial co-pro to old stuff, before nRF52 support. 1:25:55 - Demo …( using tio on arch linux ) 1:28:59 - Ryzen 16-core - speedier than a raspberry pi 4 1:31:39 - simulating the file system in a dictionary 1:33:32 book recommendations for ble ? 1:35:01 Blinka "server side" thing, to control a PiZero? Is that possible or we hit a limitation of BLE on the Blinka side? ( To have the BLE workflow for a Pi. ) 1:35:36 https://www.amazon.com/Make-Bluetooth-Projects-Raspberry-Smartphones/dp/1457187094 for Sander Vesik's question. 1:38:41 Any news about the "convergence" with BLE on micropython? /Both MP and CP were developing separately at the same time. / Low level, so blinka can build on top... to make it CP like. 1:45:34 outline for OHS 1:48:23 Soooo at 15 minutes is it a shallow dive ? 1:48:49 plan for next weeks - don’t block Trevor 1:50:12 wrap up

Tami Lane found an incredible resource in Corepower Yoga when the Seasonal Affective Disorder that comes with living in Portland led her to the hot yoga studio. She made her way to LA and into CPY management, and that's where the sanctuary shifted. Because of the high turnover, the massive growth, the lack of clear leadership and expectation, she found herself struggling with managing the CPY studio while still needing to hold down a job as a hairstylist to make enough money. Good news though: she's making her way to Ecuador, where she and her partner are hoping to offer the teachings of yoga through retreats and trainings. ___ Here's all the info on the Business Bootcamp VIP Day: https://www.splendid.yoga/businessbootcamp For more info about Morgan, check out: https://www.splendid.yoga/ Here's the Manage Your Money Yogically FB group. http://www.facebook.com/groups/splendidyoga Here's the main podcast page. http://www.splendid.yoga/mymypodcast --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app Support this podcast: https://anchor.fm/splendidyoga/support

Elyse Pancheri, mother of Eagle, Master of Crow, worked at CPY for 7 years and loved the community and the students. When I asked her what changed, it was all the things that have been recorded in lawsuits against the company, including the unpaid wages lawsuit for which the plaintiffs were awarded $1.65 million in a class action lawsuit. (A quick google search revealed two other lawsuits in which Corepower Yoga is currently embroiled: backing out of a M&A deal, and continuing to charge its members for classes after they have canceled their accounts). (Lawyers like to get involved in businesses with big investors.) You can hang out with Elyse for nerdy, body positive yoga at Hawt Mess Yoga on YouTube: https://www.youtube.com/channel/UCopJoaUymG8wGi_n0XqaVHg For more information about Morgan: https://www.splendid.yoga/ Here's the link for the Splendid Money Mastermind: https://www.splendid.yoga/mastermind Here's the Manage Your Money Yogically FB group. http://www.facebook.com/groups/splendidyoga Here's the main podcast page. http://www.splendid.yoga/mymypodcast --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app Support this podcast: https://anchor.fm/splendidyoga/support

Aliza worked for CPY for 3.5 years. She loved that the training she took made her feel equipped to teach, and the community that allowed her to look forward to her time behind the desk bonding with teachers and students. She also had to work 3 other jobs to support herself while working nearly 40 hours a week at Corepower. The toxic positivity - the "attitude of gratitude" - was used as a reason not to give her a raise for the work she was doing, and she found her requests for a more reasonable working wage fell on deaf ears, which was frustrating for a company rooted in feedback. It was also frustrating when the teacher trainings moved into quality over quantity. In order to support the expansion, CPY had to train and hire a lot of teachers - quickly. This led to a generation of teachers that were not well-educated in anatomy and how to safely direct students. Aliza now teaches online at privateyogaathome.com. You can also find her at her social media homes: Facebook.com/groups/healfromheartbreak Instagram.com/healfromheartbreak Facebook.com/AlizaPowerYoga Instagram.com/AlizaPowerYoga ___ Here's all the info on the Business Bootcamp Mastermind: https://www.splendid.yoga/mastermind For more info about Morgan, check out: https://www.splendid.yoga/ Here's the Manage Your Money Yogically FB group. http://www.facebook.com/groups/splendidyoga Here's the main podcast page. http://www.splendid.yoga/mymypodcast --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app Support this podcast: https://anchor.fm/splendidyoga/support

Kari Cowell worked for CPY for 4 years under both the franchise and the corporate studios. A bit of a nomad, she loved that she knew she could get a job teaching yoga anywhere she moved, and she had the comfort of the familiarity of corporate branding. The pay structure was frustrating, as it was for many teachers, for the level of work and experience, but ultimately, it fed Kari's desire to be her own boss. She's taken advantage of lockdown, took a bunch of business courses (including one of mine!) and now offers her work online as a spiritual healer, yoga teacher, and dating coach - yay! You can check out here stuff here: www.instagram.com/kariluyoga (wellness) www.instagram.com/kerbuhbuhbear (writing) For more information about Morgan: https://www.splendid.yoga/ Here's the link for the Splendid Money Mastermind: https://www.splendid.yoga/mastermind Here's the Manage Your Money Yogically FB group. http://www.facebook.com/groups/splendidyoga Here's the main podcast page. http://www.splendid.yoga/mymypodcast --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app Support this podcast: https://anchor.fm/splendidyoga/support

Anushree walked into the studio for her first C1 and was humbled, like so many of us are, by her first yoga class. She used the practice to heal her PCOS (Poly-cystic Ovarian Syndrome), and now works as a PCOS coach with women around the world to help heal their bodies. Coach Nush loved the management at the Atlanta Corepower Yoga where she worked, and experienced and incredible breadth of community that encouraged her to take a teacher training, try out, and coach two teacher trainings herself. One of the best parts about CPY is its ability to give students the support network through these big transitions. Here is Anushree's instagram: Www.instagram.com/coachanush For more information about Morgan: https://www.splendid.yoga/ Here's the link for the Splendid Money Mastermind: https://www.splendid.yoga/mastermind Here's the Manage Your Money Yogically FB group. http://www.facebook.com/groups/splendidyoga Here's the main podcast page. http://www.splendid.yoga/mymypodcast --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app Support this podcast: https://anchor.fm/splendidyoga/support

Tony Baker was able to use the marketing skills and community to build his own fitness business, FitGroupUSA. He eventually grew out of the model with his entrepreneurial spirit, but had a lot of gratitude for the family he was able to cultivate from the CPY community. He differentiates himself from the CPY lineage by listening to his community and acknowledging their feedback with a reason for why or why he can't take their suggestion. Like everyone I spoke with, the low pay for managers and teachers was a problem. It's a systemic problem: we as a culture don't value education. Tony's business contracts with fitness professionals at a rate 3-6 times what CPY pays its employees. They offer everything from chair yoga to qi gong to hula-hooping. Register for free as a fitness professional at: fitgroupusa.com For more information about Morgan: https://www.splendid.yoga/ Here's the link for the Splendid Money Mastermind: https://www.splendid.yoga/mastermind Here's the Manage Your Money Yogically FB group. http://www.facebook.com/groups/splendidyoga Here's the main podcast page. http://www.splendid.yoga/mymypodcast --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app Support this podcast: https://anchor.fm/splendidyoga/support

You might know Joe's voice if you're an avid gamer (he's a voiceover actor!), or you might know him from teaching at CPY. He loved the teachers, the sense of community, and the fact that he got stronger and more flexible the more he practiced there. Because he didn't rely on CPY for a primary source of income, getting paid wasn't an issue for him, but he did see it become an issue when the most popular teachers weren't incentivized to stay. The corporate yoga community in general needs to be more open to receiving and acting on the feedback it receives, and he felt the managers didn't have the authority or training to make decisions that would make the studio a better place. You can check out Joe's work at his web site: http://www.joecapvo.com/ ___ Here's all the info on the Business Bootcamp Mastermind: https://www.splendid.yoga/mastermind For more info about Morgan, check out: https://www.splendid.yoga/ Here's the Manage Your Money Yogically FB group. http://www.facebook.com/groups/splendidyoga Here's the main podcast page. http://www.splendid.yoga/mymypodcast --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app Support this podcast: https://anchor.fm/splendidyoga/support

Nicci is a Charleston, SC yoga teacher. This month she is helping to teach a Dryuary yoga challenge, 28 to stay sober and to take 20 yoga classes at CPY in Charleston, SC in the month of February. We also discuss Nicci's recent accident and how her yoga practice has helped her in her physical recovery.  

Elisabeth chats with Kristine Schumann [@sconnieyogi] about her yoga journey, what it's like to be a yoga instructor, the law of attraction, + intention setting. If you are interested in going through yoga teacher training or learning more about the law of attraction, you'll definitely want to tune in! Notes: — Kristine's CorePower Yoga schedule— Sign up to stay tuned for more info on staying involved with CHAARG post grad— CPY's teacher training— Yoga With Adrianne— Sat Nam, Kundalini in Chicago— Jess Lively's podcast, all about The Law Of Attraction— Tapas

A gift from CPY and fellow yogi Aaron English: Believe from his newest album “american [Fever] dream.” Download MP3 (right click and “Save Target As…”)

A gift from CPY and Krishna Das to you: Narayana/For Your Love from his brand new album, “Heart as Wide as the World.” Download MP3 (right click and “Save Target As…”)

Ziel dieser Arbeit sollte die Entwicklung einer automatisierten, hochparallelen und nachweis-starken Sequenzierungsmethode für Proteine im Rahmen der Proteomanalytik sein. Zur Kon-zeption der Methodik sollte dabei mit der sogenannten Leitersequenzierung eine Kopplung aus enzymatischem Aminosäureabbau und MALDI-Massenspektrometrie zum Einsatz kom-men. Die experimentellen Grundparameter für die Leitersequenzierung im Bezug auf die nasschemischen Sequenzierungsschritte und die massenspektrometrischen Messung wurden dabei im ersten Teil der Arbeit zunächst manuell ausgearbeitet. Im zweiten Abschnitt wurden dann die Möglichkeiten einer Automatisierung der so erarbeiteten Methode zur Leitersequen-zierung auf verschiedenen Ebenen evaluiert. Da der enzymatische Abbau eines Gesamtproteins durch dessen intrinsische Eigenschaften wie Sekundärstruktur, Tertiärstruktur (Zugänglichkeit der Proteintermini für die Exopepti-dase) oder Lösungsverhalten erschwert ist, wurden die Sequenzierungsprotokolle der Leiter-sequenzierung für proteolytisch erzeugte, RP-HPLC-gereinigte Spaltfragmente von Proteinen optimiert. Zudem besitzt die MALDI-MS, wie auch andere gängige massenspektrometrische Verfahren, im Massenbereich über ca. 5-10 kDa eine zu geringe Auflösung und Genauigkeit, um eine eindeutige Zuordnung der Massendifferenzen in den Leiterpeptidspektren zu den durch Exopeptidase abgespaltenen Aminosäure zu erlauben. Bei Versuchen mit verschiede-nen Endopeptidasen stellte sich – entgegen theoretischen Erwartungen – heraus, dass vor allem auch Proteinspaltungen mit Endoproteinase GluC (im Phosphatpuffer) und Chymotrypsin, neben Spaltungen mit Endoproteinase LysC, gute Voraussetzungen für die nachfolgende enzymatische Sequenzierung liefern. Mit den Proteinfragmenten aus Spaltungen der Endoproteinasen GluC (im Phosphatpuffer) und Chymotrypsin wurden bei den enzymatischen Sequenzierungen die höchsten Sequenzabdeckungen erzielt. In der technischen Ausführung wurden alle Experimente im Hinblick auf eine Sequenzierung direkt auf dem Target (on-target) optimiert. Geringfügige Anpassungen der erarbeiteten Methoden erlaubten jedoch auch eine Sequenzierung von Peptiden, die zuvor auf PVDF-Membran (Immobilon PSQ) aufgetragen wurden. Ein relativ kontrollierter und reproduzierba-rer Abbau war in einem Temperaturbereich von ca. 25-35°C möglich. Die enzymatischen Sequenzierungen erfolgten ausschließlich mit kommerziell erhältlichen Exopeptidasepräpara-tionen. Eine zusätzliche Aufreinigung der eingesetzten Enzyme erwies sich als nicht notwen-dig. Um eine einfache Interpretation der massenspektrometrischen Resultate zu gewährleisten, wurden nur Monopeptidylpeptidasen eingesetzt. Für C-terminale Sequenzierungen wurden mit CPY, CPP, CPW, CPA und CPB Carboxypeptidasen unterschiedlicher Spezifität einzeln und in verschiedenen Kombinationen untersucht. Von der N-terminal Seite aus wurden Sequenzierungen mit APM, LAP, API und AAP durchgeführt. Eine Betrachtung der Sequen-zierungen zeigt dabei, dass zumeist nur aus kombinierten Resultaten der Anwendung verschiedener Carboxy- beziehungsweise Aminopeptidasen eine ausreichende Sequenzinfor-mation erhalten werden kann. C-terminal ist der Anteil sequenzliefernder Fragmente bei der Verwendung von CPY, sowie bei den sequenziellen Peptidasenkombinationen sb(CP-I) und den Peptidasenmischungen pb(CP) mit maximal 42% am höchsten. Hier treten auch vermehrt längere Teilsequenzen mit bis zu 12 AS auf. N-terminal hebt sich APM als Einzelpeptidase mit überdurchschnittlich guten Sequenzresultaten gegenüber den anderen Aminopeptidasen ab. Bis zu 34% der getes-teten Peptide liefern allein schon mit diesem Enzym eine Sequenzinformation. Zudem wurden längere Teilsequenzen mit bis zu 9 AS im Vergleich zu anderen Aminopeptidasen häufiger erhalten. Sequenzierungen mit Aminopeptidasenmischungen bei N-terminaler Sequenzierung brachten im Gegensatz zu Carboxypeptidasenmischungen bei C-terminaler Sequenzierung kaum deutliche Vorteile. Auch sehr lange Sequenzabschnitte mit bis zu 16 AS N-terminal und bis zu 25 AS C-terminal wurden in Einzelfällen erhalten. C- und N-terminal am häufigsten wurden jedoch aus den Leiterspektren Teilsequenzen mit bis zu 6 AS erhalten (85% aller sequenzliefernden Proteinfragmente). Der größte Teil der Sequenzabdeckung stammt mit 70% aus Teilsequenzen mit 3-9 AS. Unter dem Hauptaspekt einer möglichen Steigerung der Sequenzinformation bei den Leiter-sequenzierung wurden unterschiedliche Peptidderivatisierungen untersucht. Die gezielte N-terminale Modifizierungen brachte in vielen Fällen durch die resultierende Massenverschie-bung des derivatisierten Peptids einen Gewinn an C-terminaler Sequenzinformation. Gegen-über entsprechend nicht-modifizierten Peptiden konnten durch die Massenverschiebung auch Leiterpeptide beobachtet werden, die sonst mit Signalen der MALDI-Matrix interferieren. Bei Modifikationsreagenzien, die eine fixierte positive Ladung oder ein ausgedehntes, delokali-siertes Elektronensystem ins Peptid einbrachten, wie z.B. Sulforhodamin B oder FMOC-NHS wurde dabei zusätzlich auch eine sehr gute Response im Massenspektrum beobachtet. Die Empfindlichkeiten lagen selbst bei der Sequenzierung der Rohprodukte solcher Derivate im unteren fmol-Bereich. Das charakteristische Isotopenmuster Brom-haltiger Peptidderivate erlaubte weiterhin ein stark vereinfachtes Auslesen der Leitersequenz aus dem Massenspekt-rum. Während die Leitersequenzierung allgemein keine Unterscheidung der isobaren Aminosäuren Leucin und Isoleucin erlaubt, gelang die Unterscheidung der ebenfalls isobaren Aminosäuren Lysin und Glutamin nach einer schnellen und selektiven Acetylierung des Lysins mit anschließender C-terminaler Sequenzierung.Im Hinblick auf die Analyse post-translationaler Modifikationen wurden insbesondere Phosphorylierungen eingehender untersucht. Dabei war sowohl bei der N-terminalen, als auch bei der C-terminalen Leitersequenzierung ein enzymatischer Abbau der phosphorylierten Aminosäuren zu beobachten und damit die entsprechende Phosphorylierungsstelle schnell und eindeutig zu identifizieren. Die N-terminale Sequenzierung mit APM lieferte die besten Resultate und ermöglichte sowohl den Abbau von Phosphotyrosin wie auch Phosphoserin, während der C-terminale Abbau sich auf Phosphotyrosin beschränkt zeigte. Aus der Summe der erhaltenen Resultate (Sequenzen) folgt, dass die Leitersequenzierung unter den gegebenen Voraussetzungen – insbesondere der limitierenden Verfügbarkeit zusätzlicher Exopeptidasen mit ergänzenden Spaltungsspezifitäten – im wesentlichen als Instrument zur schnellen Generierung kurzer Sequenztags geeignet ist. Auf diesem Gebiet stellt die erarbeitete Leitersequenzierung im Vergleich zur Edman-Sequenzierung eine wesentlich schnellere Alternative dar. Im Gegensatz zu rein massenspektrometrischen Sequenzierungen ist die Interpretation der Sequenzen im Massenspektrum stark vereinfacht und daher zumeist eindeutig. Die Empfindlichkeit der Methode ist stark von der untersuchten Peptidsequenz abhängig. Generelle Werte für die Empfindlichkeit lassen sich somit nicht angeben, die Resultate lassen jedoch für eine Peptidausgangsmenge im oberen fmol-Bereich (1000-500fmol) eine Leitersequenzierung ausnahmslos möglich erscheinen. Die Ergebnisse von Verdünnungsreihen zeigen zudem, dass auch nach Sequenzierung von Peptidmengen im mittleren bis unteren fmol-Bereich (50-10fmol) ein Auslesen der Peptidsequenz aus dem Massenspektrum zumeist möglich ist. Zum Erreichen solcher Empfindlichkeiten ist die weit-gehende Minimierung von Suppressionseffekten und damit die Verwendung von DHB als MALDI-Matrix eine notwendige Voraussetzung. Eine Evaluierungsstudie mit vier verschiedenen Proteinen führt zum Schluss, dass die Metho-dik auch für die de novo Sequenzierung unbekannter Proteine ein hohes Potential birgt. Die ermittelte Sequenzabdeckung der überlappenden Spaltfragmente lag bei maximal 80%. Im Bereich prolinhaltiger Sequenzabschnitte fehlen Überlappungen dabei am häufigsten, da auf Seiten der N-terminalen Sequenzierung geeignete Exopeptidasen zu Spaltung der Iminbindung nicht verfügbar waren. Zur Herstellung von Oligonucleotidsequenzen, die dann als Hybridisierungssonden in der Nucleinsäureanalytik eingesetzt werden, ist die Länge der erhaltenen Sequenzabschnitte jedoch in fast allen Fällen ausreichend. Die Methoden „MALDI-LS 1.42s“ für die Probenpräparation mit dem Pipettierroboter MultiPROBE II, sowie „multiprobe_5“ für die MALDI-MS Messung mittels AutoXecute auf dem Bruker Reflex III Massenspektrometer, erlauben eine Umsetzung der manuell ausgear-beiteten Methodik auf ein vollständig automatisiertes System. Eine Excel-Tabellenvorlage, die in konvertierter Form von beiden beteiligten Geräten gelesen werden kann, ermöglicht eine zentrale und einfache Dateneingabe für die Proben. Diese Art der Dateneingabe erlaubt im Zusammenspiel mit dem ausgearbeitetem Automatisierungspro-gramm eine vollkommen flexible, individuelle Behandlung der einzelnen Proben. Die erfor-derliche „Ortspräzision“ bei der Abgabe der Flüssigleiten auf dem MALDI-Target konnte durch eine entsprechenden ausgelegte Performance-Datei für den Pipettiervorgang erreicht werden. Querkontaminationen beim Pipettieren wurden durch organische Spülschritte in der Methode eliminiert. In der Summe lieferten die auf dem automatischen Pipettiersystem mit der Methode MALDI-LS 1.42s präparierten Proben im Massenspektrum vergleichbare Abbauspektren und somit auch die gleiche Sequenzinformation, wie entsprechend manuell präparierte Proben. Bei den automatischen MALDI-MS Messungen war eine Anpassung der Parameter insbeson-dere aufgrund der bevorzugten Verwendung der DHB-Matrix notwendig. Ein erhöhter Aceto-nitrilgehalt von 50% im Lösungsmittel der DHB sorgte für eine Verbesserung der Präparation im Bezug auf die automatische AutoXecute Messung. Mit speziellen Rasterkoordinaten, die bei der Laserabtastung der einzelnen Probenspots auf die besonderen Kristallisationseigen-schaften der DHB-Matrix zugeschnitten wurden, konnten gute Ergebnisse erzielt werden. Vorteile zeigten die DHB-Präparationen, im Vergleich zu CHCA-Präparationen, in der Toleranz gegenüber geringfügigen Mengen von Puffersalzen, wie sie bei der enzymatischen Sequenzierung üblich sind. Die Toleranzschwelle für den Abbruch der automatischen Messung musste jedoch bei den enzymatisch sequenzierten Proben und Verwendung von DHB-Matrix im Vergleich zu Messungen salzfreier Peptidproben in CHCA-Matrix erhöht werden, was im Durchschnitt zu etwas längeren Messzeiten führte. Die in dieser Arbeit weiterentwickelten Methoden zur enzymatischen on-target Sequenzie-rung von Peptiden erlauben somit, in Verbindung mit den beschriebenen Hard- und Software-komponenten zur automatischen Probenpräparation und MALDI-MS Messung, deren Einsatz für eine schnelle Sequenzierung in der Proteinanalytik. Zusätzliche Verbesserungen könnten jedoch, bei entsprechender Verfügbarkeit, noch durch Exopeptidasen mit ergänzender Spaltungsspezifität (z.B. X-Pro Aminopeptidase, EC 3.4.11.9) erzielt werden. Auf Seiten der Automatisierung ergäben sich durch die ausschließliche Ver-wendung eines 96er oder 384er Mikrotiterplattenformat auf allen Geräten (Pipettierrobot und MALDI-MS) deutliche Vereinfachungen in der Methode, bei gleichzeitig noch höherem Probendurchsatz.