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