Running Group 2 Library 3 on a Flongle to confirm successful library prep.
Need 10fmol DNA in 5uL EB. Avg. length is ~700bp, and G2L3 library is 15.3ng/uL = 35.48fmol/uL
\[ \frac{10fmol}{5uL\ EB}\ = \ \frac{25.48fmol}{17.74uL\ EB} \]
So mix 1uL G2L3 with (17.74-1) = 16.74uL EB
Flongle flow cell:
ID: AYK963
89 pores on 8/13/25 check
86 pores on today’s pore check
66 pores after loading
I’ve noticed the Flongle flow cells lose a lot of pores during loading, and community posts suggest this is a) common, and b) related to the ONT loading protocol being too harsh. This time I tried using the community-developed gentle negative pressure loading protocol for Flongles.
I still lost almost 25% of pores, but that’s better than last time (G3L1 Flongle), when I lost almost 50% of pores. I was also extra careful to seal the cover tape around all ports after loading.
Notes:
Barcode 24 is low, but that sample had a high proportion of reads in the G2L2 MinION run, so maybe this will help offset. Think its ok to run on a MinION.