Selected article for: "cdna synthesis and RNA template"
Author: Park, Donghyun; Huh, Hee Jae; Kim, Yeon Jeong; Son, Dae-Soon; Jeon, Hyo-Jeong; Im, Eu-Hyun; Kim, Jong-Won; Lee, Nam Yong; Kang, Eun-Suk; Kang, Cheol In; Chung, Doo Ryeon; Ahn, Jin-Hyun; Peck, Kyong Ran; Choi, Sun Shim; Kim, Yae-Jean; Ki, Chang-Seok; Park, Woong-Yang
Title: Analysis of intrapatient heterogeneity uncovers the microevolution of Middle East respiratory syndrome coronavirus
  • Document date: 2016_11_23
    • ID: xgp2vx6o_31
    Snippet: Five microliters of viral RNA from each sample was used as a template for cDNA synthesis using a Superscript III First-Strand Synthesis System (Life Technologies). Equal amounts of cDNA product were used to perform PCR with a Herculase II Fusion DNA polymerase (Agilent Technologies). For full-genome sequencing, primers described by Cotten et al. (2013a) were used with minor modifications for efficient amplification. A set of 60 primers was used t.....
    Document: Five microliters of viral RNA from each sample was used as a template for cDNA synthesis using a Superscript III First-Strand Synthesis System (Life Technologies). Equal amounts of cDNA product were used to perform PCR with a Herculase II Fusion DNA polymerase (Agilent Technologies). For full-genome sequencing, primers described by Cotten et al. (2013a) were used with minor modifications for efficient amplification. A set of 60 primers was used to generate fifteen 2.5-kb overlapping amplicons (four primers per amplicon) and three additional primers were added for the extreme termini of the genome (Supplemental Table S5 ). Primers used for the targeted sequencing are listed in Supplemental Table S6 . Primers and nucleotides were removed from the final PCR products using a MiniElute PCR purification kit (QIAGEN).

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