Haynes Lab:Notebook/CRISPR Editing/2015/06/27: 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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:aspirate media | :aspirate media | ||
:add | :add 400ul PBS, rock plate, aspirate PBS | ||
:add | :add 200ul trypsin, let sit for a few minutes, tap plate to unstick cells | ||
:add | :add 500ul media, pipette to collect cells, move to 1.5mL microcentrifuge tube | ||
:remove tubes from hood | :remove tubes from hood | ||
:spin in benchtop centrifuge at 0.2x1000RCF for 3 minutes | :spin in benchtop centrifuge at 0.2x1000RCF for 3 minutes | ||
:remove media with P1000 | :remove media with P1000 | ||
:resuspend in 400ul cold PBS | :resuspend in 400ul cold PBS | ||
: | :filter 200ul for flow, use remaining 200ul for gDNA extraction | ||
<br> | <br> | ||
Latest revision as of 01:02, 27 September 2017
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06/27/2015transfected gal4-eeds with puro and dox with four dilutions of g034 in triplicate. did the following twice, once for each drug treatment
Step 6: add 100ul to each well
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