IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-8-14
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*'''Insert:''' | *'''Insert:''' | ||
*#Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P | *#Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P | ||
| - | *''' | + | *'''Backbone:''' |
*#Digest J04500\S-P, \X-P (for Stage I constructs and BB high-copy) | *#Digest J04500\S-P, \X-P (for Stage I constructs and BB high-copy) | ||
*'''KaiABC ligation:''' | *'''KaiABC ligation:''' | ||
Revision as of 13:49, 14 August 2006
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To-do
- Midiprep pSB4A, J04500 (online instructions here, page 16)
- Insert:
- Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P
- Backbone:
- Digest J04500\S-P, \X-P (for Stage I constructs and BB high-copy)
- KaiABC ligation:
- KaiABC\X-P + J04500\S-P (stage I)
- KaiABC\X-P + pSB4K3 (well-formed BB)
- Sequencing:
- Sequence miniprepped KaiA/B/C + GA vectors, natural KaiA
- Western blots:
- Ligate GFP dev and pSB4A3
- Inoculate GFP dev + pSB1A2
E-gel images of colony PCR of J04500 + KaiA/B/C
