Haynes Lab:Notebook/CRISPR Editing/2015/05/27

From OpenWetWare

Jump to: navigation, search
Project name Main project page
Previous entry      Next entry

05/27/2015

Redo qPCR with the right control primers, TBP
Dilutions for 2ng gDNA per well:

Tube label Cell type gRNA plasmid 2 ug DNA ul lipo rep ng/┬ÁL dilute 1:20 ul dilution ul water
1Gal4-EEDg034-0.53181.0584.051.7314.02
2Gal4-EEDg034-0.53286.6974.331.6114.14
3Gal4-EEDg034-0.55123.891.195.869.89
4Gal4-EEDg034-0.55224.7671.245.6510.10
5Gal4-EEDg034-13185.324.271.6414.11
6Gal4-EEDg034-13270.2443.511.9913.76
7Gal4-EEDg034-15113.3870.6710.465.29
8Gal4-EEDg034-15223.2131.166.039.72
9Luc14g034GFP--1104.2395.211.3414.41
10Luc14g034GFP--264.9653.252.1613.59
11Luc14g034GFP--339.0961.953.5812.17
12Gal4-EED+doxg034GFP--116080.8814.88
13Gal4-EED+doxg034GFP--21296.451.0914.66
14Gal4-EED+doxg034GFP--31326.61.0614.69
15water--------15.75



Step 2: make primer master mixes.
Will have to add 3.5x to each triplicate gDNA tube so can calculate mastermixes from that
14 samples, 1 tube for no template control, total of 15 tubes per primer.

' 1rxn 3.5/triplicate tube 15.7
primer310.5164.85
SYBR7.526.25412.125
Total10.536.75576.975

Make these mixes for each primer pair, add 36.75 to each triplicate tube, then pipette 15ul into 3 well of the plate.


Personal tools