User:David K. Barclay/Notebook/Controlling Pancreas Cell Fate Using Transcription Factors/2014/10/27

Cell Fate Switch by Synthetic Transcription Factors

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New Primers Created from Roche Applied Science https://www.roche-applied-science.com/shop/CategoryDisplay?catalogId=10001&tab=&identifier=Universal+Probe+Library&langId=-1#tab-3 New Primers for next set of gene targets set on 10/27 from paper: "Generation of Functional Human Pancreatic Beta Cells In Vitro" (Pagliuca et al. 2014) PRIMERS LIST   Roche Name Left Primer Right Primer UPL probe INS1 NM_008386.3 cagagaggaggtactttggactataaa gccatgttgaaacaatgacct 105, cat.no. 04692241001 NKX6-1 NM_144955.2 cccggagtgatgcagagt gaacgtgggtctggtgtgtt 103, cat.no. 04692217001 MNX1 NM_019944.2 gatgccggacttcagctc agctgctggctggtgaag 60, cat.no. 04688589001 SLC30A8 NM_172816.3 gctgcttcagcaatatgcttc cagactcccagcaacgtgt 53, cat.no. 04688503001 KLF9 NM_010638.4 ctcagaactgcttttaacattaggg aacactttcctttttagctcgtg 32, cat.no. 04687655001 PCSK2 NM_008792.4 ggcgtgtttgcattagcttt gcacagtcagatgttgcatgt 85, cat.no. 04689097001 OTHER: Plasmid Verification of PcTFs Restriction Digest Table Checked plasmid minipreps with EcoRI/PstI digests Reagent Volume Expected: 1. hPCD = 1921 bp, 4592 bp 15 μL/lane; 1% agarose; Ladder DNA(plasmid) 5.0 μL 10X FD Buffer 1.5 EcoRI 1.0 PstI 1.0 dH2O 6.5   15 μL --> 37°C/ 15 min. Conclusion All bands at ~4900 and ~2000; indicating MV2 vector and PcTF insert, respectively.