IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-28: Difference between revisions
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== Western blot images == | |||
Western blot images without the annoying rectangle: | Western blot images without the annoying rectangle: | ||
[[Image:cyano_WB_CN_3.jpg|thumb]] | [[Image:cyano_WB_CN_3.jpg|thumb|left]] | ||
[[Image:cyano_WB_CN_3.tif|thumb|left]] | |||
<br style="clear:both";/> | |||
== 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. |
Revision as of 09:30, 28 October 2006
Western blot images
Western blot images without the annoying rectangle:
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.