Hi all,
I have 2 small questions concerning the 16s sequencing protocol for MiSEq:
1) Some samples have very low concentration so we are considering loading a 2pM library instead of the more standard 4pM. Anyone having bad (low) clustering doing this or no problem when using the v3 MisEq kit (2 X 300 bp) ?
2) There are various older threads discussing the PHIX % to be added to the library. The protocol says 5 % (v3 kits again), but the rep says 10-20 %. Any comments on this ?
thanks
Greg
I have 2 small questions concerning the 16s sequencing protocol for MiSEq:
1) Some samples have very low concentration so we are considering loading a 2pM library instead of the more standard 4pM. Anyone having bad (low) clustering doing this or no problem when using the v3 MisEq kit (2 X 300 bp) ?
2) There are various older threads discussing the PHIX % to be added to the library. The protocol says 5 % (v3 kits again), but the rep says 10-20 %. Any comments on this ?
thanks
Greg
Comment