I want to confirm successful library prep and compare the two “versions” of library prep for my Group 1 samples. To do this, I’ll first run a small portion of each library on Flongle flow cells.
Flongles run for ~24 hrs, so doing just Library 1 today.
Library 1 has a concentratio of 2.56 ng/uL, and an average fragment size of 1700bp (based on TapeStation). For Flongle, final library should be 5-10 fmol of DNA in 5uL EB. 5-10fmol is equivalent to 5.2ng-10.5ng of 1700bp DNA.
For Library 1, prepped Flngle final library as follows:
3uL Library 1
2uL EB
Final DNA quantity of 3uL*2.56ng/uL = 7.68ng DNA, or 7.33fmol.
Materials:
Flongle Sequencing Expansion:
REF: EXP-FSE002
LOT: FSE002.30.0005
Flongle Adapter:
SN: A100007434
Flongle Flow Cell:
LOT: 33001524
SN: B028104085
ID: AYW935
Launched MinKnow software, inserted Flongle Adapter, and ran hardware check. Then inserted Flongle Flow Cell and ran pore check: 59 pores (just above the warranty limit of 50 pores)
Followed Oxford Nanopore Technologies Native Barcoding 96 Kit protocol for Flongle flow cell loading.
After loading library, flow cell only registered 44 pores :(
Ran Flongle for 24 hrs.