Haynes Lab:Notebook/CRISPR Editing/2015/02/06

Project name

Main project page Previous entry      Next entry

02/06/2015 Trying Surveyor on g023 with a shorter transcript. Still probably too long but I know these primers work and it's still like half of the transcript I was using before. First, PCR with phusion: ' 1rxn 6.2 H20 30.8 190.96 HF 5X MasterMix 10 62 dNTPs 0.2 1.24 FP P159 2.5 15.5 RP P170 2.5 15.5 Total 46 285.2 Cell type treatment rep ng/µL ul for 200ng Luc14 g023 1 82.358 2.43 Luc14 g023 2 76.437 2.62 Luc14 g023 3 119.281 1.68 GAL4-EED +dox g023 1 122.844 1.63 GAL4-EED +dox g023 2 102.434 1.95 GAL4-EED +dox g023 3 101.862 1.96 ' 1rxn MM1 46 Template 2 DMSO 1.5 Phusion 0.5 Total 50 PCR 98°C for 3 min 30 cyles 98°C for 10s 69°C for 30s 72°C for 30s 72°C for 10 mins 4°C