Running Group 1, Library 4 on a Flongle to confirm successful library prep.
Need 10fmol DNA in 5uL EB. Assume 1700bp average length: 10fmol = 10.5ng DNA.
Library 4 is 36.6ng/uL, so too concentrated to accurately pipette 10.5ng. Instead, make a 4X mix:
\(\frac{10.5ng\ DNA}{5uL\ EB} = \frac{42ng\ DNA}{20uL\ EB}\)
To get volume of G1L4 to add:
\(\frac{42ng\ DNA}{36.6 ng/uL\ G1L4} = 1.15uL\ G1L4\)
So added 1.15 uL G1L4 to (20-1.15uL) = 18.85uL EB. Then used 5uL of this to make Flongle loading library.
Flongle flow cell
SN: B028104167
LOT: 33001524
ID: AYL248
60 pores on check
33 pores after loading
This is a pretty significant drop in pores after library loading, even for a Flongle.
Ran for 24 hours.
This Flongle run had very poor results – low output and quickly degrading pores. After discussion with Dan, though, he thinks this was a problem with the flow cell itself (supported by the dramatic pore loss after loading), not the library. I’ll go ahead with the Group 1 Library 4 prep and try it on a MinION.