User:Cassandra M Barrett/Notebook/Haynes Lab/2015/08/25: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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== | ==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 | |||
Latest revision as of 01:08, 27 September 2017
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T2A-EGFP fragment and MV11 plasmid DigestPurpose: 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
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