Haynes Lab:Notebook/CRISPR Editing/2015/11/08: Difference between revisions

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(Autocreate 2015/11/08 Entry for Haynes_Lab:Notebook/CRISPR_Editing)
 
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==03//2015==
==11/08/2015==
 
Nested PCR of g032 Luc14 samples (last run had a band in the negative ctrl lane)<br>Ran on gel.


<br><br>
Cut 100ng of each HDR sample with EcoRI. <br>
Controls
:Donor PCR products
:Unedited Luc14 gDNA PCR w P197/198
:No enzyme ctrl for each<br>
Total of 9 with enzyme and 9 without
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Cell type'''
| align="center" style="background:#f0f0f0;"|'''gRNA'''
| align="center" style="background:#f0f0f0;"|'''donor'''
| align="center" style="background:#f0f0f0;"|'''rep'''
| align="center" style="background:#f0f0f0;"|'''ng/µL'''
| align="center" style="background:#f0f0f0;"|'''ul for 100ng'''
| align="center" style="background:#f0f0f0;"|'''water'''
|-
| Luc14||g031||g031 EcoRI||1||35.13||2.85||1.15
|-
| Luc14||g031||g031 EcoRI||2||65.872||1.52||2.48
|-
| Luc14||g031||g031 EcoRI||3||58.161||1.72||2.28
|-
| N/A||N/A||g031 EcoRI||N/A||63.7||1.57||2.43
|-
| Luc14||g034||g034 EcoRI||1||36.115||2.77||1.23
|-
| Luc14||g034||g034 EcoRI||2||28.507||3.51||0.49
|-
| Luc14||g034||g034 EcoRI||3||37.64||2.66||1.34
|-
| N/A||N/A||g034 EcoRI||N/A||132||0.76||3.24
|-
| Luc14||N/A||N/A||N/A||121||0.83||3.17
|}
Incubate at 37°C for 15 minutes, heat inactivate at 80°C for 5 minutes. <br>
Dilute 1:10, run on BA.


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Revision as of 07:07, 8 November 2015

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11/08/2015

Nested PCR of g032 Luc14 samples (last run had a band in the negative ctrl lane)
Ran on gel.



Cut 100ng of each HDR sample with EcoRI.
Controls

Donor PCR products
Unedited Luc14 gDNA PCR w P197/198
No enzyme ctrl for each

Total of 9 with enzyme and 9 without

Cell type gRNA donor rep ng/µL ul for 100ng water
Luc14 g031 g031 EcoRI 1 35.13 2.85 1.15
Luc14 g031 g031 EcoRI 2 65.872 1.52 2.48
Luc14 g031 g031 EcoRI 3 58.161 1.72 2.28
N/A N/A g031 EcoRI N/A 63.7 1.57 2.43
Luc14 g034 g034 EcoRI 1 36.115 2.77 1.23
Luc14 g034 g034 EcoRI 2 28.507 3.51 0.49
Luc14 g034 g034 EcoRI 3 37.64 2.66 1.34
N/A N/A g034 EcoRI N/A 132 0.76 3.24
Luc14 N/A N/A N/A 121 0.83 3.17

Incubate at 37°C for 15 minutes, heat inactivate at 80°C for 5 minutes.
Dilute 1:10, run on BA.


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