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

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==To-do==
==To-do==
-
*#Midiprep pSB4A, J04500
+
*'''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 13:33, 14 August 2006




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

Click for legend
Click for legend
Click for legend
Click for legend


It appears that our vector self-ligated.

To-do

  • Midiprep pSB4A, J04500
  • 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
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