Dinh/NOTES/2010-2-1
Jump to navigation
Jump to search
NEBNext DNA Sample Prep Master Mix Set 1
Purpose: To prepare sample for next-generation sequencing and for preparation of expression libraries.
1. NEBNext End Repair Module
Recommended use: 1-5ug DNA fragments (100-1000bp) in <= 85ul
Reaction Mix:
1-85ul Fragmented DNA 10ul NEBNext End Repair Reaction buffer (10x) 5ul NEBNext Ennd Repair Enzyme Mix (thaw on ice) variable DNAse/RNase free, sterile water -------------------- 1x volume = 100ul
Program: incubate for 30min at 20C
Purification: use column or beads and elute with Elution Buffer to volume of 37ul
2. NEBNext dA-tailing Module
Reaction Mix
37ul end-repaired, blunt DNA 5ul NEBNext dA-tailing reaction buffer 3ul Klenow Fragment (3'->5' exo) variable DNAse/RNase free, sterile water