Team:Caltech/Week 6

Monday 7/19

• Group meeting!
  • Discussed issues with cloning and next week's focus.
• Performed a digest (now for 2hr) and ligations (L18-23). Transformed into DH5α.
• Turns out that the sequencing primers we ordered (G00100 & G00101) are incorrect - they appear to bind on the wrong sides of the insert. We ordered new primers designed to bind to the BioBrick prefix and suffix.
• Ran gel of digest product to verify correct bands.

Digest test gel with US and DS digested bricks. (7/19)

Tuesday 7/20

• All ligation transformations failed. Will try 16hr ligations and purifying digest products with a PCR purification kit.
• Made new growth curves testing AIBN effect on cells in LB
• Performed another live/dead assay
  • Had solutions of cells in glycerol and AIBN
  • Plated cells every hour for three hours
  • Combined glycerol solutions with LB and plated the solution

Wednesday 7/21

• Ran another digest:

Thursday 7/22

• Ran gel of digest products from yesterday to verify correctness of the digestion.

Digest test gel with upstream, downstream, and backbone digested bricks. (7/22)

Friday 7/23

Weekend 7/24-25