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

K562 RNA submission to DNASU

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DNASU Submission I'll be submitting six samples to DNASU today: K562 24hr no plasmid K562 24hr KAH126-MV2 rep1 K562 24hr KAH126-MV2 rep2 K562 48hr no plasmid K562 48hr KAH126-MV2 rep1 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