Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/21: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Line 49: | Line 49: | ||
<center> | <center> | ||
Results of the PCR | Results of the PCR products, with H2B on the left and LOV on the right. The gel was loaded with 5 μL samples. | ||
</center> | </center> | ||
Line 57: | Line 57: | ||
'''Assemblies''' | '''Assemblies''' | ||
# | # H2B - X/S - 410 bp + pSB1A3 - X/S - 2000 bp | ||
* Digests: | |||
** In a 30 μL reaction, cut H2B with XbaI and SpeI | |||
** In a 30 μL reaction, cut LOV with XbaI and SpeI | |||
{| class="wikitable" border=1 cellpadding="5" cellspacing="0" | |||
|- | |||
| Reagent || H2B PCR || LOV PCR | |||
|- | |||
| DNA || 8.0 || 8.0 | |||
|- | |||
| XbaI || 1.0|| 1.0 | |||
|- | |||
| SpeI || 1.0 || 1.0 | |||
|- | |||
| 10x buffer || 3.0 || 3.0 | |||
|- | |||
| dH<sub>2</sub>O || 10.0 || 10.0 | |||
|- | |||
| Total vol. || 30. 0 || 30.0 | |||
|} | |||
* Run entire 30 μL on a 1% gel (use the big fat-tooth well comb) | |||
* Cut band out of gel (use UV box) | |||
<center> | |||
[[Image:VN_Gel_11-14-12.png|200px|Results of from the digests are shown.]] | |||
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]] | |||
</center> | |||
<center> | |||
# H2B = 410 | |||
# LOV = 461 | |||
</center> | |||
'''Dephosphorylation''' | |||
'''Ligation''' | '''Ligation''' |
Revision as of 14:04, 21 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> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
11/21/12
PCR program:
Results of the PCR products, with H2B on the left and LOV on the right. The gel was loaded with 5 μL samples.
Assemblies
Dephosphorylation
Ligation
Proceed directly to Transformation. See http://openwetware.org/wiki/Haynes:Assembly101
|