Dinh/NOTES/2010-2-1

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Revision as of 01:04, 31 January 2010 by >Dinh (Created page with '=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…')
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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
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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