Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/06/17

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K562 RNA submission to DNASU Main project page
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DNASU Submission

I'll be submitting six samples to DNASU today:

  1. K562 24hr no plasmid
  2. K562 24hr KAH126-MV2 rep1
  3. K562 24hr KAH126-MV2 rep2
  4. K562 48hr no plasmid
  5. K562 48hr KAH126-MV2 rep1
  6. K562 48hr KAH126-MV2 rep2

Setup for sample prep:

Sample ID Conc. (ng/µL) 10x dilution Vol. for 100ng (µL) Vol. H2O (µL)
K562 24hr no plasmid969.
K562 24hr KAH126-MV2 rep1609.460.91.68.4
K562 24hr KAH126-MV2 rep2670.767.11.58.5
K562 48hr no plasmid802.
K562 48hr KAH126-MV2 rep1970.797.11.09.0
K562 48hr KAH126-MV2 rep2835.483.51.28.8

cDNA Synthesis

I'll be making two 2 µg aliquots of each sample (including pMaxGFP controls) for cDNA synthesis, for a total of 16 reactions.

Setup for cDNA synthesis:

Sample ID Conc. (ng/µL) Vol. for 2 µg (µL) Vol. primer (µL) vol. dNTP (µL) vol. H2O (µL)
K562 24hr no plasmid969.22.1115.9
K562 24hr KAH126-MV2 rep1609.43.3114.7
K562 24hr KAH126-MV2 rep2670.73.0115.0
K562 48hr no plasmid802.22.5115.5
K562 48hr KAH126-MV2 rep1970.72.1115.9
K562 48hr KAH126-MV2 rep2835.42.4115.6
K562 24hr pMaxGFP638.03.1114.9
K562 48hr pMaxGFP802.22.4115.6

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