User:Karmella Haynes/Notebook/PcTF Genomics/2015/06/25: Difference between revisions
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* Final H3#_MV11 builds verified on 6/24/15 | * Final H3#_MV11 builds verified on 6/24/15 | ||
* Co-transfect H3#_MV11 with pSP-guide RNA plasmid (built by Ben Nyer) | * Co-transfect H3#_MV11 with pSP-guide RNA plasmid (built by Ben Nyer) | ||
* Use Luc14 cells (from Rene) | * Use HEK293 Luc14 cells (from Rene) | ||
# H340_MV11 + | # H340_MV11 + g034_pSPgRNA | ||
# H360_MV11 + | # H340_MV11 (no gRNA) | ||
# H380_MV11 + | # H360_MV11 + g034_pSPgRNA | ||
# | # H360_MV11 (no gRNA) | ||
# H380_MV11 + g034_pSPgRNA | |||
# H380_MV11 (no gRNA) | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Trfxn. table --> | {| class="wikitable" border="0" cellspacing="3" <!-- Trfxn. table --> | ||
|-valign="top" | |-valign="top" | ||
| <u>Well</u> || <u>H3#_MV11 (1.0 μg)</u> || <u> | | <u>Well</u> || <u>H3#_MV11 (1.0 μg)</u> || <u>g034_pSPgRNA (1.0 μg)</u> || <u>dH<sub>2</sub>O</u> || <u>OptiMEM</u> || <u>PLUS rgt.</u> || <u>Lipo LTX</u> | ||
|- | |- | ||
| 1 || | | 1 || 5.0 || 1.6 || 13.4 || 570.0 || 2.5 || 7.5 | ||
|- | |- | ||
| 2 || | | 2 || 5.0 || --- || 15.0 || 570.0 || 2.5 || 7.5 | ||
|- | |- | ||
| 3 || | | 3 || 5.0 || 1.6 || 13.4 || 570.0 || 2.5 || 7.5 | ||
|- | |- | ||
| 4 || | | 4 || 5.0 || --- || 15.0 || 570.0 || 2.5 || 7.5 | ||
|- | |- | ||
| 5 || | | 5 || 5.6 || 1.6 || 12.8 || 570.0 || 2.5 || 7.5 | ||
|- | |- | ||
| 6 || | | 6 || 5.6 || --- || 14.4 || 570.0 || 2.5 || 7.5 | ||
|} | |} | ||
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* Add PLUS --> 5 min./ RT | * Add PLUS --> 5 min./ RT | ||
* Add Lipo LTX --> 30 min./ RT | * Add Lipo LTX --> 30 min./ RT | ||
* | * Cells - refresh growth medium w/ ps-free DMEM | ||
* Add 600 μL complexes | * Add 600 μL complexes, drop-wise | ||
* Incubate overnight | * Incubate overnight | ||
Revision as of 18:22, 25 June 2015
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06/25/15
Cas-tone - transfections
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