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

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[[Image:VN_Gel_11-14-12.png|Results of from the digests are shown.]]
[[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 enzyme digests are shown, with H2B on the left, LOV in the middle, and pSB1A3 on the right. The gel was loaded with 30 μL samples.
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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Revision as of 04:45, 14 November 2012

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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.

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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