Download Agilent SureGuide gRNA Synthesis Kit Guide

Agilent SureGuide gRNA Synthesis Kit
Protocol
Agilent SureGuide gRNA Synthesis Kit
Kit Components and Storage Conditions 4
Required and Optional Equipment and Reagents 5
Introduction to gRNA Synthesis 6
Preprotocol Considerations 7
CRISPR and tracr sequences 7
DNA template requirements and design 8
T7 RNA polymerase promoter primer 9
Template input 10
Procedures 12
Transcribe the gRNA from the DNA template 12
Purify the gRNA 13
gRNA Analysis 15
Spectrophotometric analysis 15
Electrophoretic analysis 15
Supplementary Protocols 17
Fill-in procedure for “Template option B” 17
Purification of double stranded template 18
Supplementary Design Information 19
gRNA and DNA template sequences for an example DNA target 19
References 23
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