Haynes Lab:Notebook/CRISPR Editing/2014/10/27: Difference between revisions

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(Autocreate 2014/10/27 Entry for Haynes_Lab:Notebook/CRISPR_Editing)
 
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==07//2014==
==10/27/2014==
 
Phosphorylated and annealed gRNAs 23, 25, 31-38 for  cloning into pX330.


{| {{table}}
|-
|1 ul oligo 1 (100µM)
|-
|1 ul oligo 2 (100µM)
|-
|1 ul 10X T4 Ligation Buffer (NEB)
|-
|6.5 ul ddH2O
|-
|0.5 ul T4 PNK (NEB)
|-
|10 ul total
|}


Anneal in a thermocycler using the following parameters:
:37°C 30 min
:95°C 5 min 
:ramp down to 25°C at 5°C/min
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Revision as of 12:54, 27 October 2014

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10/27/2014

Phosphorylated and annealed gRNAs 23, 25, 31-38 for cloning into pX330.

1 ul oligo 1 (100µM)
1 ul oligo 2 (100µM)
1 ul 10X T4 Ligation Buffer (NEB)
6.5 ul ddH2O
0.5 ul T4 PNK (NEB)
10 ul total

Anneal in a thermocycler using the following parameters:

37°C 30 min
95°C 5 min
ramp down to 25°C at 5°C/min


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