Haynes Lab:Notebook/CRISPR Editing/2015/03/06

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03/06/2015

miniprep of pMax GFP 6.2 from Lonza and KAH60

plasmid colony 260 280 260/280 ng/µL ul for 05.ng
pMax GFP 6.210.5520.2991.848551.7790.906159894
pMax GFP 6.220.3480.1871.861347.5531.438629504
KAH6010.1020.0561.817101.8064.911301888
KAH6020.2780.1481.878278.4111.795906053


plasmid ng/µL ul for 05.ng g044 403.212 1.240042459 g031 288.187 1.734984576

plated cells look great, all wells are about 90% confluent. Will proceed with all 48 transfections

KAH60 transfections


Step 1 ' ' ' '
Making 1ug/10ul plasmid stocks
ng/ulul plasmidul H2O
KAH60278.41151.0091.00
pMax GFP 6.2551.77914.8610222675.13897774
g032452.73913.4748.53
g031288.18721.1740.83
g025235.9525.8536.15
g044403.21215.1346.87
Step 2
make DNA + optimem mixes
KAH60add 554 ul optimem
pMax GFP 6.2add 320 ul optimem
g032add 242 ul optimem
g031add 242 ul optimem
g025add 242 ul optimem
g044add 242 ul optimem
Step 3
add plus reagent to DNA+ opti
KAH60add 14.2ul plus
pMax GFP 6.2add 8.2ul plus
g032add 6.2ul plus
g031add 6.2ul plus
g025add 6.2ul plus
g044add 6.2ul plus
step 4
Make optimem + lipo mastermix
Optimem2265.4
lipo144.6
Step 5
mix dna mix and lipo mix
KAH60add 710 lipo mix
pMax GFP 6.2add 410 lipo mix
g032add 310 lipo mix
g031add 310 lipo mix
g025add 310 lipo mix
g044add 310 lipo mix
Step 6
add 100ul dropwise to each well


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