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

Gel from 7/10/07