IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-8-14: Difference between revisions
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==To-do== | ==To-do== | ||
*'''Midiprep''' pSB4A, J04500 (online instructions [http://www1.qiagen.com/literature/handbooks/PDF/PlasmidDNAPurification/PLS_Plasmid/1034637_HB_QIAGENPlasmid_112005.pdf | *'''Midiprep''' pSB4A, J04500 (online instructions [http://www1.qiagen.com/literature/handbooks/PDF/PlasmidDNAPurification/PLS_Plasmid/1034637_HB_QIAGENPlasmid_112005.pdf here], page 16) | ||
*'''Insert:''' | *'''Insert:''' | ||
*#Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P | *#Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P |
Revision as of 10:48, 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
- 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
E-gel images of colony PCR of J04500 + KaiA/B/C
It appears that our vector self-ligated.