Haynes Lab:Notebook/CRISPR Editing/2016/05/31

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05/31/2016

qPCR for ChIP
Plate 2
Dropping Primer set 3 because the input calls were around 39 so there's no chance we'd see any pulldown DNA. Josh gave me new primers against GAPDH promoter he's been using for his ChIPs. Will test those for one input set on this plate.

Also adding wells with high variability between technical replicates. All the Luc14 samples on this plate are re-runs.

Primer set 1 MM, 39 wells

' 1 well 42
2x SYBR mix 7.5 315
750nmol F/R primer mix 3 126
PCR water 2.5 105
total 13 546


Primer set 2 MM, 33 wells

' 1 well 36
2x SYBR mix 7.5 270
750nmol F/R primer mix 3 108
PCR water 2.5 90
total 13 468

Primer set 3 MM, 12 wells

' 1 well 14
2x SYBR mix 7.5 105
750nmol F/R primer mix 3 42
PCR water 2.5 35
total 13 182

16.05.30 qPCR Plate 2 screen shot.png



Plate 3

Primer set 1 MM, 21 wells

' 1 well 24
2x SYBR mix 7.5 180
750nmol F/R primer mix 3 72
PCR water 2.5 60
total 13 312


Primer set 2 MM, 57 wells

' 1 well 60
2x SYBR mix 7.5 450
750nmol F/R primer mix 3 180
PCR water 2.5 150
total 13 780


16.05.30 qPCR Plate 3 screen shot.png




Plate 4
Run by Josh Cutts, the best
Primer set 3 MM, 84 wells

' 1 well 87
2x SYBR mix 7.5 652.5
750nmol F/R primer mix 3 261
PCR water 2.5 217.5
total 13 1131


13ul master mix into each well, 2ul of sample into each well, following guide below:

16.05.31 qPCR Plate 4 screen shot.png