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

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.pngK562 RNA submission to DNASU Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

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 plasmid 969.2 96.9 1.0 9.0
K562 24hr KAH126-MV2 rep1 609.4 60.9 1.6 8.4
K562 24hr KAH126-MV2 rep2 670.7 67.1 1.5 8.5
K562 48hr no plasmid 802.2 80.2 1.2 8.8
K562 48hr KAH126-MV2 rep1 970.7 97.1 1.0 9.0
K562 48hr KAH126-MV2 rep2 835.4 83.5 1.2 8.8

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 plasmid 969.2 2.1 1 1 5.9
K562 24hr KAH126-MV2 rep1 609.4 3.3 1 1 4.7
K562 24hr KAH126-MV2 rep2 670.7 3.0 1 1 5.0
K562 48hr no plasmid 802.2 2.5 1 1 5.5
K562 48hr KAH126-MV2 rep1 970.7 2.1 1 1 5.9
K562 48hr KAH126-MV2 rep2 835.4 2.4 1 1 5.6
K562 24hr pMaxGFP 638.0 3.1 1 1 4.9
K562 48hr pMaxGFP 802.2 2.4 1 1 5.6