User:Cassandra M Barrett/Notebook/Haynes Lab/2015/08/25

From OpenWetWare
Jump to navigationJump to search
Project name Main project page
Previous entry      Next entry

T2A-EGFP fragment and MV11 plasmid Digest

Purpose: to prepare the T2A-EGFP insert and MV11 vector for cloning via double restriction digest

Both fragment and backbone run using same digest protocol

Methods:

Backbone/fragment 5uL XbaI 1uL Kpn2I 1uL FD Buffer 1uL Water 2uL

10 min at 37C in block Heat inactivate: 80C for 5 min, 65C for 20 min

Both samples cleaned up using Qiagen PCR clean up kit and eluted in 30uL of Elution Solution