User:Karmella Haynes/Notebook/miR Trigger/2009/12/21

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12/21/09

 * &#x2713; Order oligos for RT-PCR analysis of YFP(miRNA) vs. RFP expression
 * &#x2713; Split cells: U2OS plain (1x 1:10 flask); JDS33-47 U2OS (1x 1:10 flask, 2x 6-well ab-free 2.5x105/well, 1x 24-well ab-free)
 * &#x2713; Cultures for minipreps: KAH93, 94, 95/MV2 (2 each)

Order oligos > For RT-PCR analysis of YFP(from transgene JDS33/MV2) and RFP(from tansgenes KAH93, 94, 95/MV2) expression; 100 bp amplicons > Design MV2 vector primers to detect any DNA contamination (non-transcribed sequence) in cDNA samples
 * 1) YFP_RTPCR f1: 5'-GCGCGCCTGAGAACTTCAGG (B-globin exon 2)
 * 2) YFP_RTPCR r1: 5'-ctcgcccttgctcactctag (scar-YFP1)
 * 3) YFP_RTPCR f2: 5'-gcatggacgagctgtacaag (YFP1 end)
 * 4) YFP_RTPCR r2: 5'-tggccgtttacgtcgccgtc (YFP2 near start)
 * 5) RFP_RTPCR f1: 5'-gcctctaacattgagacagc (Gal4DB near end)
 * 6) RFP_RTPCR r1: 5'-cggaaattccggcgatacag (Gal4DB end)
 * 7) RFP_RTPCR f2: 5'-gcatggacgagctgtacaag (RFP1 near end)
 * 8) RFP_RTPCR r2: 5'-gagccctccatgtgcaccttg (RFP2 near start)
 * 9) MV2_RTPCR f:  5'-CGACGCTCAAGTCAGAGGTG (backbone, non-coding)
 * 10) MV2_RTPCR r:  5'-GCAGGGTCGGAACAGGAGAG (backbone, non-coding)


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