Selected article for: "cdna synthesis and RNA template"
Author: Schwarz, Megan C.; Sourisseau, Marion; Espino, Michael M.; Gray, Essanna S.; Chambers, Matthew T.; Tortorella, Domenico; Evans, Matthew J.
Title: Rescue of the 1947 Zika Virus Prototype Strain with a Cytomegalovirus Promoter-Driven cDNA Clone
  • Document date: 2016_9_28
    • ID: 0i1abjfa_30
    Snippet: RT-PCR analysis of splicing. RNA was extracted from cells 48 h after transfection with the wild-type MR766 plasmid or Pol(Ϫ) plasmid or infection with the parental MR766 virus or a mock control using the PureLink RNA minikit (Thermo Fisher Scientific, Waltham, MA). The extracted RNA was used as a template for random hexamer-primed cDNA synthesis using the SuperScript III First-Strand synthesis system (Thermo Fisher Scientific, Waltham, MA). Five.....
    Document: RT-PCR analysis of splicing. RNA was extracted from cells 48 h after transfection with the wild-type MR766 plasmid or Pol(Ϫ) plasmid or infection with the parental MR766 virus or a mock control using the PureLink RNA minikit (Thermo Fisher Scientific, Waltham, MA). The extracted RNA was used as a template for random hexamer-primed cDNA synthesis using the SuperScript III First-Strand synthesis system (Thermo Fisher Scientific, Waltham, MA). Five hundred nanograms of cDNA was used for PCR using the Expand High-Fidelity PCR system (Roche Life Sciences, Indianapolis, IN) with oligonucleotides flanking the region into which the intron was cloned: ME-O-1722 (ATGTCCGCTTGAGCACAGAG) and ME-O-1738 (AGCGATGTTGTCAGTGCGTG). PCR products were ligated into pGEM-T vector (Promega, Madison, WI) for subsequent propagation in bacteria and sequencing.

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