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 mix7.5315
750nmol F/R primer mix3126
PCR water2.5105
total13546


Primer set 2 MM, 33 wells

' 1 well 36
2x SYBR mix7.5270
750nmol F/R primer mix3108
PCR water2.590
total13468

Primer set 3 MM, 12 wells

' 1 well 14
2x SYBR mix7.5105
750nmol F/R primer mix342
PCR water2.535
total13182

Image:16.05.30 qPCR Plate 2 screen shot.png



Plate 3

Primer set 1 MM, 21 wells

' 1 well 24
2x SYBR mix7.5180
750nmol F/R primer mix372
PCR water2.560
total13312


Primer set 2 MM, 57 wells

' 1 well 60
2x SYBR mix7.5450
750nmol F/R primer mix3180
PCR water2.5150
total13780


Image: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 mix7.5652.5
750nmol F/R primer mix3261
PCR water2.5217.5
total131131


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

Image:16.05.31 qPCR Plate 4 screen shot.png


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