Haynes Lab:Notebook/CRISPR Editing/2014/12/09: Difference between revisions

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==12/09/2014==
==12/09/2014==
PCR of gDNA for screening with Surveyor. Will use P197 and P198 and Taq MM.  
'''PCR of gRNA treated Luc14 gDNA for screening with Surveyor'''
Annealing temp 53°C, ext time 2min, 10min final ext. 30 cycles
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''ul per 1 rxn'''
|-
| 2x Taq MM||25
|-
| P197 100uM||2.5
|-
| P198 100uM||2.5
|-
| Sample 100ng/ul||2
|-
| H2O||18
|-
| Total||50
|}
<br>
<br>
50ul reactions, 200ng gDNA, 2.5ul of 100uM primers.
:95°C for 3 min
:30 cycles
::95°C for 30 s
::53°C for 30 s
::72°C for 2 min
:72°C for 10 min
<br>
 


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Revision as of 10:25, 10 December 2014

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12/09/2014

PCR of gRNA treated Luc14 gDNA for screening with Surveyor

' ul per 1 rxn
2x Taq MM 25
P197 100uM 2.5
P198 100uM 2.5
Sample 100ng/ul 2
H2O 18
Total 50


95°C for 3 min
30 cycles
95°C for 30 s
53°C for 30 s
72°C for 2 min
72°C for 10 min




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