Difference between revisions of "IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-8-14"

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m (Moved the to-do list up.)
(To-do)
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==To-do==
 
==To-do==
*'''Midiprep''' pSB4A, J04500
+
*'''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 10:47, 14 August 2006

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To-do

  • Midiprep pSB4A, J04500 (online instructions [1], page 16)
  • Insert:
    1. Digest miniprepped KaiA/B/C in GeneArt plasmid with X-P
  • bkb:
    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 + pSB4K3 (well-formed BB)
  • 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


It appears that our vector self-ligated.