User:Karmella Haynes/Notebook/PcTF Genomics/2015/06/27: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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==mm/dd/yy==
==06/27/15==
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* Cas-tone - cell transfections (Gal4EED/luc)
* Cas-tone - cell transfections (Gal4EED/luc)

Latest revision as of 01:02, 27 September 2017

Pc-TF Genomics Main project page
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06/27/15

  • Cas-tone - cell transfections (Gal4EED/luc)


Cas-tone - transfections

  • Final H3#_MV11 builds verified on 6/24/15
  • Co-transfect H3#_MV11 with pSP-guide RNA plasmid (built by Ben Nyer)
  • Use HEK293 Gal4EED/luc cells, no dox (from Rene)
  1. H340_MV11 + g034_pSPgRNA
  2. H340_MV11 (no gRNA)
  3. H360_MV11 + g034_pSPgRNA
  4. H360_MV11 (no gRNA)
  5. H380_MV11 + g034_pSPgRNA
  6. H380_MV11 (no gRNA)
Well H3#_MV11 (1.0 μg) g034_pSPgRNA (1.0 μg) dH2O OptiMEM PLUS rgt. Lipo LTX
1 5.0 1.6 13.4 570.0 2.5 7.5
2 5.0 --- 15.0 570.0 2.5 7.5
3 5.0 1.6 13.4 570.0 2.5 7.5
4 5.0 --- 15.0 570.0 2.5 7.5
5 5.6 1.6 12.8 570.0 2.5 7.5
6 5.6 --- 14.4 570.0 2.5 7.5
  • Pre-warm ps-free DMEM
  • Make 20 μL plasmid/dH2O mixes in sterile 1.5 ml tubes
  • Add OptiMEM
  • Add PLUS --> 5 min./ RT
  • Add Lipo LTX --> 30 min./ RT
  • Cells - refresh growth medium w/ ps-free DMEM
  • Add 600 μL complexes, drop-wise
  • Incubate overnight



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