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

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(→November 9, 2012)
(→November 9, 2012)
Line 43: Line 43:
<center>
<center>
-
[[Image:Gel_11-9-12.png|Results of the PCR product digests are shown, with H2B on the left and LOV on the right.]]
+
[[Image:VN_Gel_11-14-12.png|Results of from the digests are shown.]]
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]]
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]]
</center>
</center>
<center>
<center>
-
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.
+
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.
</center>
</center>

Revision as of 07:44, 14 November 2012

Project name Main project page
Previous entry      Next entry

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 from the digests are shown. Image:KAH_Fermentas_GeneRuler_1kbplus.jpg

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.


Retrieved from "http://www.openwetware.org/wiki/Haynes_Lab:Notebook/Investigating_Photo-Switchable_Synthetic_Nucleosomes/2012/11/09"
Personal tools