IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-8-14

To-do

• Midiprep pSB4A, J04500 (online instructions here, page 16)
• Insert:
  1. Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P
• Backbone:
  1. Digest J04500\S-P, \X-P (for Stage I constructs and BB high-copy)
• KaiABC ligation:
  1. KaiABC\X-P + J04500\S-P (stage I)
  2. KaiABC\X-P + pSB1AK3 (well-formed BB)
  3. KaiABC\X-P + B0034\S-P (stage I)
• Sequencing:
  1. Sequence miniprepped KaiA/B/C + GA vectors, natural KaiA
• Western blots:
  1. Ligate GFP dev and pSB4A3
  2. Inoculate GFP dev + pSB1A2

E-gel images of colony PCR of J04500 + KaiA/B/C

Click for legend

Click for legend