Haynes Lab:Notebook/CRISPR Editing/2015/02/22

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02/22/2015

Testing qPCR primers with dilution curves using Luc14 g023 treated rep 1 gDNA (82.4ng/ul)

' For Rev Target
Primer set 1P207P208pX330
Primer set 2P209P210pX330
Primer set 3P211P212pX330
Primer set 4P201P202GAPDH
Primer set 5P203P204GAPDH
Primer set 6P205P206MMP12

Primer mix

Reagent Single well x6.5
2x SYBR Green Master Mix7.548.75
750 nM F/R primer mix319.5
Total vol.10.568.25

Template mixes
Make 1:10 dilution of gDNA sample so the new concentration is 8.24ul/ul.

Dilution name Make dilution stock Make template MM
1:1 diladd 1.21ul to 8.79ul water6.5 ul dil to 22.75 H2O
1:10 diladd 1 ul to 9 ul water6.5 ul dil to 22.75 H2O
1:100 diladd 1 ul to 9 ul water6.5 ul dil to 22.75 H2O
1:1000 diladd 1 ul to 9 ul water6.5 ul dil to 22.75 H2O
1:10000 diladd 1 ul to 9 ul water6.5 ul dil to 22.75 H2O
Water only29.25 H2O
1:11:101:1001:10001:10000No temp ctrl
P set 1
P set 2
P set 3
P set 4
P set 5
P set 6


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