Materials needed

Reagent Producer Catalog #
Mung Bean Nuclease NEB M0250S
SalI-HF NEB R3138T
Agencourt AMPure XP Beckman Coulter A63881

Procedure

  1. Setup a restriction digest using 1 ug of DSN library
Component Volume
10X Cutsmart Buffer 4 ul
SalI-HF Enzyme (100 units) 1 ul
Molecular Grade H20 22.75 ul
DSN Enriched Library 12.25 ul
Total Volume 40 ul

  1. Incubate reactions in thermocycler at 37°C for at least 8 hours, prefereably 12-16 hours.

  2. Using the same tubes from the previous step combine the following:

Component Volume
Digested Library 40 ul
10X Mung Bean Nuclease buffer 4.5 ul
Mung Bean Nuclease (10 units per ul) 0.5 ul
Total Volume 45 ul
  1. Incubate at 30°C for 30 minutes
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