User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/26: Difference between revisions
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* ChIP-seq: use input results to determine enrichment of U2OS H3K27me3 signals with Seqman NGen | * ChIP-seq: use input results to determine enrichment of U2OS H3K27me3 signals with Seqman NGen | ||
* TRIzol prep: U2OS PcTF+ & mock cells | |||
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* Set control inputU2OS as "yes" for "Is control?" | * Set control inputU2OS as "yes" for "Is control?" | ||
* Set inputU2OS as control for 6_aln_sorted. | * Set inputU2OS as control for 6_aln_sorted. | ||
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-- | '''U2OS TRIzol prep'''<br> | ||
# +PcTF, plate 1-A | |||
# +PcTF, plate 1-B | |||
# mock, plate 4-A | |||
# mock, plate 4-B | |||
(Should provide more than enough RNA. Discarded other cultures) | |||
* Cells were 100% confluent | |||
* Discarded growth medium | |||
* Added 2 mL TRIzol directly to plates | |||
* Incubated at r.t. for ~5 min. | |||
* Collected lysed cells from plate with gentle scraping (pipette tip) | |||
* Transferred 500 μL aliquots to 2.0 mL tubes. Stored at -80°C | |||
** 8x PcTF+ samples | |||
** 8x mock samples | |||
Revision as of 20:16, 26 June 2013
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06/26/13
ChIP seq analysis with Input control Array Star analysis - general instructions
H3K27me3 enrichment in U2OS
U2OS TRIzol prep
(Should provide more than enough RNA. Discarded other cultures)
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