Hi Annie, when you next run samples, you will make up flush buffer and load that into the buffer port. This will flush all of the storage buffer into the waste channel.
Hi Kevin, that depends how long you run it for and what you are doing. If you are sequencing a small part of the genome, or only need to know species level (e.g. metagenomics), then you can run for a short period (~ a few hours) and get the info you need, then wash the flow cell and reuse. However, if you need detail down to SNP level for WGS, and/or you're running multiple samples, to get the detail you need you'll probably need to run it for the full 48-72 hours, so there'll be no point washing it because there won't be many pores left. Once you wash the flow cell, you should QC it again immediately and it will tell you how many active pores you've got left to work with. In theory one flow cell can be used multiple times, but you need to consider data quality and application!
Hi Maurine, it doesn't really matter, all that will happen if you add more buffer and the waste reaches the port, is that it will just bubble out of it. Equally, removing the waste shouldn't affect the ASIC panel section of the flow cell, so you can remove it if you wish
Hi Luis. The flow cells will just click into place (of course you need to make sure they are the correct way round, to ensure the electronics on the bottom fit), but we recommend being gentle with it! Once MinKNOW has displayed the number of active pores, you can then remove the flow cell (it's easiest to do so by gently lifting the waste port end up and then slowly wiggling the flow cell out - you can see Linzy do this in the video). There are no locking mechanisms in place on either the flow cell or MinION (it just has small magnets to hold the lid down)
@@pandora-id-netconsortium1846did QC 1432 pores on the flowcel. How to unplug safely the minION? I tried to eject ....but windows response is "device running" so i am closing software and then turning off the computer. Then unplug. As i get ready for my first run. Flushing is done while placed on the the minION ? Looking forward
Hi there, Should we remove storage buffer before priming the flow cell for next run?
Quite helpful and informative.
You made mention of being able to make the A and B buffers, do you where I can find information on how to make them?
these are commercial buffers that come with flow cell wash kit
Brilliant video
How to remove storage buffer before next use?
Hi Annie, when you next run samples, you will make up flush buffer and load that into the buffer port. This will flush all of the storage buffer into the waste channel.
How many times can I use one cell flow?
Hi Kevin, that depends how long you run it for and what you are doing. If you are sequencing a small part of the genome, or only need to know species level (e.g. metagenomics), then you can run for a short period (~ a few hours) and get the info you need, then wash the flow cell and reuse. However, if you need detail down to SNP level for WGS, and/or you're running multiple samples, to get the detail you need you'll probably need to run it for the full 48-72 hours, so there'll be no point washing it because there won't be many pores left. Once you wash the flow cell, you should QC it again immediately and it will tell you how many active pores you've got left to work with. In theory one flow cell can be used multiple times, but you need to consider data quality and application!
How much waste should be removed from the waste port before washing?
Hi Maurine, it doesn't really matter, all that will happen if you add more buffer and the waste reaches the port, is that it will just bubble out of it. Equally, removing the waste shouldn't affect the ASIC panel section of the flow cell, so you can remove it if you wish
test the storage buffer within the device ;)
few Q need a proper how to turn on an off the equipment. say you have done your calibration check. now what just pull the tests cell out?
Hi Luis. The flow cells will just click into place (of course you need to make sure they are the correct way round, to ensure the electronics on the bottom fit), but we recommend being gentle with it! Once MinKNOW has displayed the number of active pores, you can then remove the flow cell (it's easiest to do so by gently lifting the waste port end up and then slowly wiggling the flow cell out - you can see Linzy do this in the video). There are no locking mechanisms in place on either the flow cell or MinION (it just has small magnets to hold the lid down)
@@pandora-id-netconsortium1846did QC 1432 pores on the flowcel. How to unplug safely the minION? I tried to eject ....but windows response is "device running" so i am closing software and then turning off the computer. Then unplug.
As i get ready for my first run. Flushing is done while placed on the the minION ?
Looking forward