[Belinda]

Lambda control experiment (SQK-LSK109)

Hello everyone, I am new in using nanopore too. I just bought the starter kit for SQK-LSK109 and I have done the lambda control experiment. I am not sure why the pore is inactivated after i added my sample on the flow cell. I already checked the pore of my flow cell and it is functioning well. is there anyone why this will happen? I had followed every steps carefully in the protocol.
Thank you very much!

[Belinda]

Lambda control experiment (SQK-LSK109)

Hello everyone, I am new in using nanopore too. I just bought the starter kit for SQK-LSK109 and I have done the lambda control experiment. I am not sure why the pore is inactivated after i added my sample on the flow cell. I already checked the pore of my flow cell and it is functioning well. is there anyone why this will happen? I had followed every steps carefully in the protocol.
Thank you very much!

[jhalpin]

Hi everyone! We've been dabbling with nanopore off and on for a little over a year. Recently, I've been trying the Ligation sequencing kit with native barcoding on a version 9 flowcell. The DNA looks good going in (on a 1% gel, no fragment analyzer available) and we started with usheared 2ug and 7 samples. I changed the cleanups to 0.6 and of course the last pool cleanup is 0.5x. We use wide bore pipette tips, minimize pipetting, tap to mix, no vortex, etc. And my run predicted N50 is 54 bases. BASES. It's 15 hours into 48.. there is at least one read >600kb so there are some long reads but most are tragically short.

I'm at a loss as to what else I can try here.. any ideas? I got good yield after each cleanup so I'm inclined to think that something is happening after pooling but before/during loading on the flow cell that is shearing the DNA.


ETA we did not have this problem with the LSK109 kit alone. Adding the barcodes seemed to change something.

[Cassandra Botil]

This is interesting
How you all put out their experiences
Makes me want to read more