Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/14: Difference between revisions
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| pSB1A3|| 0.004 || 0.001 || 2.857 || 4.233 | | pSB1A3|| 0.004 || 0.001 || 2.857 || 4.233 | ||
|} | |||
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*'''[[User:Karmella Haynes|---Karmella]] 21:34, 15 November 2012 (EST)''': Next step - ligation & transformation<br> | |||
Refer to this page for a full description: http://openwetware.org/wiki/Haynes:Assembly101 | |||
* Use 20 ng of vector for each ligation = 5 μL pSB1A3 | |||
* Use 2x moles of H2B insert compared to vector: xμL H2B insert = 2 * 410 bp H2B / 2000 bp pSB1A3 * 20 ng pSB1A3 / 6.384 ng/μL H2B = '''1.28 μL''' | |||
{| class="wikitable" width=400px align="right" | |||
| || H2B || LOV || Negative Control | |||
|- | |||
| Insert DNA (X ng) || 1.28 μL || '''none''' | |||
|- | |||
| Vector DNA (50 ng) || 5.0 || 5.0 || same | |||
|- | |||
| 2x Roche Rapid Ligation buffer || 5.0 μl || same | |||
|- | |||
| New England Biolabs T4 ligase || 1.0 μl || same | |||
|- | |||
| dH<sub>2</sub>O || ___ μL || ___ μL + ''Insert'' μL | |||
|- | |||
| || 10.0 μL total || same | |||
|- | |||
| colspan="3" | Mix the reaction(s) thoroughly by flicking the tube.<br>Incubate at room temperature for 10 minutes. | |||
|} | |} | ||
Revision as of 19:34, 15 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> | ||||||||||||||||||||||||||||||||||||||||||||||
November 14, 2012
Refer to this page for a full description: http://openwetware.org/wiki/Haynes:Assembly101
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