Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/23: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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Proceed directly to Transformation. See http://openwetware.org/wiki/Haynes:Assembly101 | Proceed directly to Transformation. See http://openwetware.org/wiki/Haynes:Assembly101 | ||
[[Image:VN_H2B_and_LOV_Transformation_Trial_2.png|thumb|frame|center|There were no cultures seen the next day.]] | |||
'''Digestion''' | '''Digestion''' | ||
* Check ligations from | * Check ligations from 11/20/12 to see if any colonies contained the insert | ||
* Used 4 colonies from H2B, 4 colonies from LOV plates | * Used 4 colonies from H2B, 4 colonies from LOV plates | ||
* Each colony liquid culture followed the digestion table accordingly | * Each colony liquid culture followed the digestion table accordingly | ||
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| EcoRI || 1.0|| 0 | | EcoRI || 1.0|| 0 | ||
|- | |- | ||
| PstI || 0 || 1.0 |- | | PstI || 0 || 1.0 | ||
|- | |||
| 10x buffer || 1.5 || 1.5 | | 10x buffer || 1.5 || 1.5 | ||
|- | |||
| dH<sub>2</sub>O || 7.5 || 7.5 | | dH<sub>2</sub>O || 7.5 || 7.5 | ||
|- | |||
| Total vol. || 15.0 || 15.0 | | Total vol. || 15.0 || 15.0 | ||
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|} | |} | ||
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<center> | <center> | ||
[[Image:VN_Gel_11-23-12.png | [[Image:VN_Gel_11-23-12.png|Results of from the digests are shown.]] | ||
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]] | [[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]] | ||
</center> | </center> |
Latest revision as of 22:19, 26 September 2017
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November 23, 2012Dephosphorylation (11/23/12) > Dephosphorylation (Roche)
Proceed directly to Transformation. See http://openwetware.org/wiki/Haynes:Assembly101 Digestion
Results of the enzyme digests are shown, with four H2B bands on the left, and three LOV bands on the right. The gel was loaded with 30 μL samples. The bands are at 2000 bp, confirming the vector self ligated and there is no insert.
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