Haynes Lab:Notebook/CRISPR Editing/2015/06/24

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transfected luc14s with four dilutions of g034 in triplicate

Step 1: Diluting the DNA

' ug of DNA for 3.2 wells ul plasmid water (32ul total)
g034 0.250.8
g034 0.51.6
g034 0.752.4
g034 1.03.2

Step 2: make DNA + optimem mixes

' 1 well 3.2 x
g034 0.2539add 124.8ul optimem
g034 0.539add 124.8ul optimem
g034 0.7539add 124.8ul optimem
g034 1.039add 124.8ul optimem

Step 3: add plus reagent to DNA+ opti

' 1 well 3.2 x
g034 0.251add 3.2ul plus reagent
g034 0.51add 3.2ul plus reagent
g034 0.751add 3.2ul plus reagent
g034 1.01add 3.2ul plus reagent

Step 4: Make optimem + lipo mastermix

' 1 well for 12 wells (x 13.2)

Step 5: mix dna mix and lipo mix

g034 0.25add 160 lipo mix
g034 0.5add 160 lipo mix
g034 0.75add 160 lipo mix
g034 1.0add 160 lipo mix

Step 6: add 100ul to each well

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