Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/11/30: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Today's project is...</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Today's project is...</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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Thermal cycler program:<br> | |||
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'''Transformation''' | |||
Follow the instructions in the protocol described [http://openwetware.org/wiki/Haynes:TransformationPlasmids#Traditional_Method:_Chemically_competent_cells_.2B_ligation here]. Setting up three plates: | |||
# Sample | |||
# Negative control - pJET vector only | |||
# negative control - no plasmid | |||
==Notes== | |||
May have to perform a PCR clean-up on the sample next time before transforming cells. | |||
==Colony PCR Results== | |||
Performed colony PCR on the one colony on the sample plate. No product. Giving up on LCR and switching to Golden Gate assembly. | |||
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Latest revision as of 01:23, 27 September 2017
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OverviewPerforming LCR on the PNK-treated parts from last week. Methods
Thermal cycler program:
Transformation Follow the instructions in the protocol described here. Setting up three plates:
NotesMay have to perform a PCR clean-up on the sample next time before transforming cells. Colony PCR ResultsPerformed colony PCR on the one colony on the sample plate. No product. Giving up on LCR and switching to Golden Gate assembly. |