Dinh/NOTES/2010-2-1: Difference between revisions

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>Dinh
m (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…')
 
>Dinh
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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
--------------------
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

Latest revision as of 18:43, 10 February 2010