IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-8-14: Difference between revisions
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==To-do== | ==To-do== | ||
* | *'''Midiprep''' pSB4A, J04500 | ||
*Insert: | *'''Insert:''' | ||
*#Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P | *#Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P | ||
*bkb: | *'''bkb:''' | ||
*#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:''' | ||
*#KaiABC\X-P + J04500\S-P (stage I) | *#KaiABC\X-P + J04500\S-P (stage I) | ||
*#KaiABC\X-P + pSB4K3 (well-formed BB) | *#KaiABC\X-P + pSB4K3 (well-formed BB) | ||
*Sequencing: | *'''Sequencing:''' | ||
*#Sequence miniprepped KaiA/B/C + GA vectors, natural KaiA | *#Sequence miniprepped KaiA/B/C + GA vectors, natural KaiA | ||
*Western blots: | *'''Western blots:''' | ||
*#Ligate GFP dev and pSB4A3 | *#Ligate GFP dev and pSB4A3 | ||
*#Inoculate GFP dev + pSB1A2 | *#Inoculate GFP dev + pSB1A2 |
Revision as of 10:33, 14 August 2006
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E-gel images of colony PCR of J04500 + KaiA/B/C
It appears that our vector self-ligated.
To-do
- Midiprep pSB4A, J04500
- Insert:
- Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P
- bkb:
- 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