Jessica Karen Wong/Notebook/2007-7-17
From OpenWetWare
Jump to navigationJump to search
Sequencing
Made glycerols, minipreped, and sent for sequencing w/ VF and VR
- E0240-1AK3 Colonies 1 and 2
- I2055-1AK3 #4 and #16
- E0240-3K3 #9
- I2057-3K3 #2 and #4
- I2055-3K3 #3
- and 2 of Jason's 507 and 528
Also sent T9002-3K3 #12 for seq with VF, VR, and internal primers
PCR
- Made 8 cell suspensions of I2055-3K3 and ran 10ul rxns of them w/ taq and a 1:30 ext time
- PCR'ed I2056 to scar and add in promoter
- Diluted I2056_R0040_F to 40 ul
- Also used I2055_R
- 1 100ul rxn at 53 w/ vent
- PCRed E0240 portion of T9002 1 100ul rxn w/ vent at 54
- Set up overnight BB PCR of E0240-3K3
- 1 100ul w/ vent at 53.5, 2:30 ext time
F2620
- PCR cleaned digests of F2620 (M/N), 1AK3 (E/P), 3K3 (E/P)
- Ligated F2620-1AK3 and F2620-3K3
- Transformed 3ul of each ligation and plated
Gels
- Ran gel of overnight gradient pcr of T9002_E0240 (gel 1) - looked good
- Ran gel of today's pcr's
- Loaded: Lad, Sp, I2056, Sp, I2055-3K3 (colonies 1-8)
- Made an overnight of Colony #1 to sequence