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Brown/21 June 2009
From 2009.igem.org
Team3
Tasks for today: • Run digest again, test E and S on tet • Genomic purification of DH5α then PCR OmpR, Run gel • PCR Taz1, Run gels • Calculate competency of gels
1) Digest of Tet (from 7/16 minipreps) with EcoRI and SpeI Concentration (ng/μL ) μL DNA μL water Tet Steph 129.3 7.737 8.27 Tet Ash 126.1 7.93 8.07 Tet Michael 71.8 13.93 2.07
Incubate till 12:00 pm (1.5 hours) Insert gel here Lane1: 1 kb ladder Lane 2: Ash Lane 3: Ash 2 Lane 4: 1 kb ladder Lane 5: Steph Lane 6: Steph 2 Lane 7: Michael Lane 8: 1kb
E, S cut successfully for Tet→ enzymes are ok
2) Genomic purification performed. Primers: OmpR BB Reverse, OmpR BB Forward Mastermix: 47 μL; Primers: 1 each, DNA: 1 μL Insert gel here Lane1: 1 kb ladder Lane 2: 1 kb ladder Lane 3: Tube 1 Lane 4: Tube 2 Lane 5: Tube 3 Lane 6: Tube 4 Lane 7: 100 bp ladder Lane 8: 1kb
→ Do not use tubes 2 and 3 of genomic DNA 3) PCR of Taz 1 Insert gel pictures here A: amplification. B: Biobrick; Numbers correspond to Taz samples from minipreps of 7/17) Gel 1 Lane 1: 1kb Lane 2: 1A Lane 3: 1B Lane 4: 2A Lane 5: 2B Lane 6: 3A Lane 7: 3B Lane 8: 1 kb
Gel 2: Lane 1: 1kb Lane 2: 4A Lane 3: 4B Lane 4: 5A Lane 5: 5B Lane 6: 6A Lane 7: 6B Lane 8: 1 kb
