- Видео 6
- Просмотров 76 166
PIROON JENJAROENPUN
Добавлен 26 ноя 2008
Flongle priming and loading #nanopore
Taylor and Max gonna show you how to do priming and loading a “Flongle” flow Cell
• 1st, insert the flongle adaptor into MinION
• Next, insert the Flongle flow cell
• Make sure to align the contact pad on the flow cell and adapter
• Then, Lift up the sticky tab and peel back the seal
• Stick the tab to avoid it folds back
• now we are ready to load the priming mix
• This step is critical, you have to make sure there is no air at the bottom of pipette tip.
• Loading Air bubble can damage flow cell
• Insert tip into sample port and slowly pipette priming buffer
• Now, load you sample. Again, no air bubble
• insert tip into sample port and slowly pipette library
• Final steps sealing the sample port ...
• 1st, insert the flongle adaptor into MinION
• Next, insert the Flongle flow cell
• Make sure to align the contact pad on the flow cell and adapter
• Then, Lift up the sticky tab and peel back the seal
• Stick the tab to avoid it folds back
• now we are ready to load the priming mix
• This step is critical, you have to make sure there is no air at the bottom of pipette tip.
• Loading Air bubble can damage flow cell
• Insert tip into sample port and slowly pipette priming buffer
• Now, load you sample. Again, no air bubble
• insert tip into sample port and slowly pipette library
• Final steps sealing the sample port ...
Просмотров: 11 167
Видео
Washing MinION Flow Cell
Просмотров 23 тыс.6 лет назад
PRECAUTION Before you start, to know more about "waste port" please watch this link first ruclips.net/video/n5P0Q2ItaPg/видео.html Created by Thidathip Wongsurawat #thidathip
Elephant Dung Project using MinION
Просмотров 1,7 тыс.7 лет назад
Without bringing DNA sample back to the lab in US. It is possible to collect fresh sample, perform DNA extraction, sequence long DNA and run metagenomic analysis. We could generate data to our collaborator within a few day before flying back. ; ) #OneMoreFlowCell Acknowledgement To my team (Gam and Niramol), your help has made my job very much easier and more fun. I really appreciate your time ...
Education Matters Biomedical Informatics
Просмотров 4267 лет назад
Education Matters Biomedical Informatics
Loading a Oxford Nanopore flow cell
Просмотров 38 тыс.7 лет назад
Loading a Flow cell in 3 steps 1. check for small bubble under the cover. Draw back a small volume to remove any bubble (a few µls) 2. Prime the Flow Cell a. Load 800 µl of the priming mix into the Flow Cell via the priming port, avoiding the introduction of air bubbles, and wait five minutes. b. Gently lift the SpotON sample port cover to make the SpotON sample port accessible. c. Load 200 µl ...
Has anyone ever used electronic pipettes, because that is what our lab has and we are having problems? We don't know how fast to make it move out to not damage the pores. Any help would be very very appreciated. Thank you!
wooow you do it realy bad .... used nanopore for 3 year and you are doing it bad .
I think overall okay. Probably should not load all in the tip to avoid introducing bubble. The flow rate can also be lower.
Hello.. I am interested in this product for research purposes but I have a question… can this product be used without the dna library? What if I want to analyze a sample from an unknown organism? What do you recommend?
Currently, the product requires the DNA library preparation step. You can extract DNA and perform Nanopore sequencing. The organism name is then determined by analyzing the DNA sequences with WIMP (EPI2ME).
How many times can the flow cell be used when cleaned properly?
We've washed flow cells no more than five times. Some of our collaborators, however, can do it more than five times. It all depends on how much yield you need and how many active pores remain after each run.
what happens if you forget to wash a flowcell and leave it out overnight? is it still reusable? (asking for a friend)
Your friend can count how many active pores there are. If the activate pores > 600, he/she may wash and reuse the flowcell. I would, however, use the washed flowcell for testing purposes rather than production.
How much priming buffer and library required
120 uL priming buffer and about 30 uL sample
Hi, can I use the video for training course?
Of course, please use it.
วันนี้ inspire มากๆเลยค่ะ 🤩 ขอขอบคุณนะคะ/ ได้ฟังที่ CRA _ ติดตามงานของอาจารย์ทั้งสองคนน้าค่ะ
ขอบคุณมากๆ เลยค่ะ พี่มีกำลังใจทำงานมากขึ้นเลยค่า
Yo thank you so much for the video. I couldnt find anything to read on how to load the sample into the Flongle flow cell, how did you find this? Also, I read about the sample lib prep to be half-volumes, do you know where to find this?
you can find it in Nanopore website > nanopore community > protocol > Genomic DNA by Ligation (SQK-LSK109)> download PDF
@@thidathipw2953 Thanks Tip! I've been using the protocols from the store, not the community section -_-
@@zarulsaurus7551 You are welcome. Enjoy using Flongle : )
Are you guys running a different protocol from this one? cdn.livechat-static.com/api/file/lc/att/7281641/1a541cf808f28569cd82b60bd43b531c/flow-cell-wash-kit-protocol-WFC_9088_v1_revD_18Sep2019-any.pdf
We used the previous version (EXP-WSH002) not the protocol you share.
I have problems with the software please help me
You can download the program from nanopore community web site. They have instruction "how to install and use the program in detail. community.nanoporetech.com/protocols/experiment-companion-minknow/v/mke_1013_v1_revam_11apr2016
@iam Nobody depends on the sample you use. For high purity DNA, we can reuse 2-3 times. Using high modified DNA (e.g.methylation), maybe just one time.
@iam Nobody Yes. My lab usually buys 48 flow cells batch which allows us to get a cheaper price per flow cell.
please I need information how to get the software and how to install the software
I remember that the protocol says drawing the waste after adding solution A and S. Maybe the protocol has been updated?
We remove the waste before and after adding the solution. Sometimes we reused a flow cell by adding A and B. If we do not remove the waste before washing again, then, we cannot add solution S. Because it's full of liquid in the waste area.
I could not find any procedure for drawing the waste before adding solution A. On current version (WKE_1012_v1_revN_08Apr2016) of protocol mentioned that remove all buffer from the waste section after you added 500 ul of Storage Buffer. So, to reuse flowcell (keep at 4C), we probably have to follow; 1) Open Priming port, Add150 ul of Solution A. 2) wait for 10 min. 3) Add 500 ul of Storage Buffer. 4) Close Priming port, remove all buffer from the waste port. 5) stored at 4-8C".
@@Matsu9Matsu I'm also confused. The protocol didn't say anything about removing any solution before adding buffers. But it seems we do need to remove waste solutions before adding buffers to avoid any overflow.
cool