Haynes Lab:Notebook/CRISPR Editing/2015/01/07: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Rene M Davis (talk | contribs) (Autocreate 2015/01/07 Entry for Haynes_Lab:Notebook/CRISPR_Editing) |
Rene M Davis (talk | contribs) |
||
Line 6: | Line 6: | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
==07/ | ==01/07/2015== | ||
Set up 2-50μL PCR reactions for untreated Luc14 gDNA and a subset of treated Luc14.<br> | |||
<br> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Mastermix 1''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
|- | |||
| ||1rxn||28.3rxns | |||
|- | |||
| H20||30||849 | |||
|- | |||
| HF 5X MasterMix||10||283 | |||
|- | |||
| dNTPs||1||28.3 | |||
|- | |||
| FP||2.5||70.75 | |||
|- | |||
| RP||2.5||70.75 | |||
|- | |||
| ||46||1301.8 | |||
|- | |||
| Mastermix 1|||| | |||
|- | |||
| ||1rxn||2.1rxns | |||
|- | |||
| MM1||46||96.6 | |||
|- | |||
| Template||2||4.2 | |||
|- | |||
| DMSO||1.5||3.15 | |||
|- | |||
| Phusion||0.5||1.05 | |||
|- | |||
| ||50||205 | |||
|} | |||
Transfer 50μL of MM2 into each of 4-PCR tubes.<br> | |||
PCR | |||
:98°C for 3 min | |||
:36 cycles | |||
::98°C for 10s | |||
::66°C for 30s | |||
::72°C for 60s | |||
:72°C for 10 min | |||
:4°C forever | |||
<br> | |||
'''Sample g039 may have too much gDNA, samples g045 and g046 were in non-autoclaved tubes, sample g046 did not have 50ul for each tube''' | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> |
Revision as of 16:20, 7 January 2015
Project name | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||
01/07/2015Set up 2-50μL PCR reactions for untreated Luc14 gDNA and a subset of treated Luc14.
Transfer 50μL of MM2 into each of 4-PCR tubes.
|