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

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2012/11/09 Entry for Haynes_Lab:Notebook/Investigating_Photo-Switchable_Synthetic_Nucleosomes)
 
(14 intermediate revisions by the same user not shown)
Line 6: Line 6:
| colspan="2"|
| colspan="2"|
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==Entry title==
==November 9, 2012==
* Insert content here...
 
* First, resuspend dry primer oligos (in blue-capped tubes) to 100 μM
* Then make working solutions of 10 μM
* Reactions:
 
# H2B-GFP plasmid + BB_H2B fwd + BB_H2B rev
# LOV plasmid + BB_LOV fwd + BB_LOV rev
 
{| class="wikitable" border=1 cellpadding="5" cellspacing="0"
|-
| 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
|-
| dH<sub>2</sub>O || 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 ∞
 
{| class="wikitable" border=0
|-
| <u>Expected:</u><br>1. H2B = 410 <br>2. LOV = 461
| <br>
|}
 
<center>
[[Image:Gel_11-9-12.png|Results of the digests are shown.]]
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]]
</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.
</center>
 




Revision as of 04:45, 14 November 2012

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Investigating_Photo-Switchable_Synthetic_Nucleosomes/2012/11/09&oldid=654431"