IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-28

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== Western blot images ==
Western blot images without the annoying rectangle:
Western blot images without the annoying rectangle:
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[[Image:cyano_WB_CN_3.jpg|thumb]]
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[[Image:cyano_WB_CN_3.jpg|thumb|left]]
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[[Image:cyano_WB_CN_3.tif|thumb|left]]
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<br style="clear:both";/>
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[[Image:cyano_WB_CN_3.tif|thumb]]
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== Miniprep of J36335 (KaiA + KaiC), J36832 (KaiB) ==
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We performed 8 minipreps each of J36335 and J36832, eluted in 30 µL EB.
 +
 
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== Digest of J36335\S-P (KaiA + KaiC), J36832\X-P (KaiB) ==
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We did 8 digests each of J36335\S-P and J36832\X-P.
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=== Reaction ===
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J36335\S-P
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* 7.6 µL DNA
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* 1 µL 10x BSA
 +
* 1 µL NEBuffer 2
 +
* 0.2 µL SpeI
 +
* 0.2 µL PstI
 +
 
 +
J36832\X-P
 +
* 7.6 µL DNA
 +
* 1 µL 10x BSA
 +
* 1 µL NEBuffer 3
 +
* 0.2 µL XbaI
 +
* 0.2 µL PstI
 +
 
 +
=== Master mixes ===
 +
J36335\S-P
 +
* 76 µL DNA
 +
* 10 µL 10x BSA
 +
* 10 µL NEBuffer 2
 +
* 2 µL SpeI
 +
* 2 µL PstI
 +
Aliquot 10µL into each tube.
 +
 
 +
J36832\X-P
 +
* 76 µL DNA
 +
* 10 µL 10x BSA
 +
* 10 µL NEBuffer 3
 +
* 2 µL XbaI
 +
* 2 µL PstI
 +
Aliquot 10µL into each tube.

Revision as of 11:30, 28 October 2006

Contents

Western blot images

Western blot images without the annoying rectangle:

Image:Cyano WB CN 3.tif

Miniprep of J36335 (KaiA + KaiC), J36832 (KaiB)

We performed 8 minipreps each of J36335 and J36832, eluted in 30 µL EB.

Digest of J36335\S-P (KaiA + KaiC), J36832\X-P (KaiB)

We did 8 digests each of J36335\S-P and J36832\X-P.

Reaction

J36335\S-P

  • 7.6 µL DNA
  • 1 µL 10x BSA
  • 1 µL NEBuffer 2
  • 0.2 µL SpeI
  • 0.2 µL PstI

J36832\X-P

  • 7.6 µL DNA
  • 1 µL 10x BSA
  • 1 µL NEBuffer 3
  • 0.2 µL XbaI
  • 0.2 µL PstI

Master mixes

J36335\S-P

  • 76 µL DNA
  • 10 µL 10x BSA
  • 10 µL NEBuffer 2
  • 2 µL SpeI
  • 2 µL PstI

Aliquot 10µL into each tube.

J36832\X-P

  • 76 µL DNA
  • 10 µL 10x BSA
  • 10 µL NEBuffer 3
  • 2 µL XbaI
  • 2 µL PstI

Aliquot 10µL into each tube.

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