Brown/21 June 2009

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