Jessica Karen Wong/Notebook/2007-7-17

Sequencing
Made glycerols, minipreped, and sent for sequencing w/ VF and VR Also sent T9002-3K3 #12 for seq with VF, VR, and internal primers
 * 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

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