Difference between revisions of "Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/09"

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* 72°C, 3 min.
 
* 72°C, 3 min.
 
* 4°C ∞
 
* 4°C ∞
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| <u>Expected:</u><br>1. H2B = 410 <br>2. LOV = 461
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[[Image:VN_Gel_11-9-12.png|150px|Results of the digests are shown.]]
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[[Image:Gel_11-9-12.png|Results of the digests are shown.]]
 
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]]
 
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]]
 
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Results of the PCR product digests are shown, with H2B on the left and LOV on the right. The gel was loaded with 5 μL samples.
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Latest revision as of 22:14, 26 September 2017

Owwnotebook icon.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

November 9, 2012

  • First, resuspend dry primer oligos (in blue-capped tubes) to 100 μM
  • Then make working solutions of 10 μM
  • Reactions:
  1. H2B-GFP plasmid + BB_H2B fwd + BB_H2B rev
  2. LOV plasmid + BB_LOV fwd + BB_LOV rev
Reagents H2B LOV
Plasmid DNA 0.5 μL 0.5
primer 1 (10 μM) 1.0 1.0
primer 2 (10 μM) 1.0 1.0
2x GoTaq mix 12.5 12.5
dH2O 10.5 10.5

PCR program:

  • 95°C, 3 min.
  • [95°C, 30 sec; 57°C, 30 sec.; 72°C, 30 sec.] x35 cycles
  • 72°C, 3 min.
  • 4°C ∞
Expected:
1. H2B = 410
2. LOV = 461

Results of the digests are shown. KAH Fermentas GeneRuler 1kbplus.jpg

Results of the PCR product digests are shown, with H2B on the left and LOV on the right. The gel was loaded with 5 μL samples.