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

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06/24/2015

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 0.25 0.8
g034 0.5 1.6
g034 0.75 2.4
g034 1.0 3.2


Step 2: make DNA + optimem mixes

' 1 well 3.2 x
g034 0.25 39 add 124.8ul optimem
g034 0.5 39 add 124.8ul optimem
g034 0.75 39 add 124.8ul optimem
g034 1.0 39 add 124.8ul optimem


Step 3: add plus reagent to DNA+ opti

' 1 well 3.2 x
g034 0.25 1 add 3.2ul plus reagent
g034 0.5 1 add 3.2ul plus reagent
g034 0.75 1 add 3.2ul plus reagent
g034 1.0 1 add 3.2ul plus reagent


Step 4: Make optimem + lipo mastermix

' 1 well for 12 wells (x 13.2)
optimem 47 620.4
lipo 3 39.6


Step 5: mix dna mix and lipo mix

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

Step 6: add 100ul to each well