Jessica Karen Wong/Notebook/2007-7-10

I2057

 * Heat shocked overnight digest of I2057 with Mfe1/Nsi1
 * PCR cleaned I2057 digest
 * Ligated with both 1AK3 and 3K3
 * Transformed 3ul of each and plated on Kan

I2056

 * PCR cleaned overnight scarring PCR of I2056
 * Ran gel of I2056 and overnight colony PCR's
 * Nothing showed up the right size
 * Got sequencing in for I2056
 * Sequence only matched plasmid, none of the part was present
 * Discarded cleaned PCR
 * Rebuilding part: ligated R0040-J04650-1AT3, R0040-J04650-1AC3
 * Transformed 3ul of each and plated on appropriate antibiotic
 * Made new Chlor plates, running low

T9002

 * Ligated T9002-3K3 and transformed
 * Also spun down rest of T9002-1AK3 transformation and plated

E0240

 * Redid BioBricking PCR of E0240-1AK3 on a gradient
 * 12 10ul PCR's from 51c to 57c
 * Used Taq and too short elongation time 1:45
 * This product was not the right size (rightmost lanes)
 * Has some at 3kb and some at around 10kb, but should be 4kb
 * Redid BB PCR overnight on same gradient with Vent and proper 2:40 elongation time
 * Made new overnight of E0240-1AK3 b/c were running out of DNA
 * Religated and transformed E0240-3K3

I2055

 * Scarred product in 1AK3 had 5 colonies on the transformation plate
 * Colony PCR-ed all 5
 * Ran a gel and nothing showed up again (the first five lanes after the ladder)
 * Did a positive control with P1010 from the registry to make sure we're doing colony PCR right
 * 1 10ul with VF and VR and extension time 1:30