Selected article for: "forward human and reverse primer"
Author: Liu, Yuan-yuan; Chen, Liang-jun; Zhong, Yan; Shen, Meng-xin; Ma, Nian; Liu, Bing-yu; Luo, Fan; Hou, Wei; Yang, Zhan-qiu; Xiong, Hai-rong
Title: Specific interference shRNA-expressing plasmids inhibit Hantaan virus infection in vitro and in vivo
  • Document date: 2016_3_14
    • ID: pwlybr2h_22
    Snippet: qRT-PCR analysis Total RNAs were extracted from the cell or tissues using TRIzol reagents (Invitrogen, USA) following the manufacturer's instructions. First-strand cDNA was synthesized by reverse transcription of 500 ng of total RNA using random primers (Promega) and M-MLV reverse transcriptase (Promega) at 25 ºC for 10 min, 37 ºC for 60 min, 85 ºC for 5 min and 4 ºC for 4 min. The amplification was performed using the following primer sets: .....
    Document: qRT-PCR analysis Total RNAs were extracted from the cell or tissues using TRIzol reagents (Invitrogen, USA) following the manufacturer's instructions. First-strand cDNA was synthesized by reverse transcription of 500 ng of total RNA using random primers (Promega) and M-MLV reverse transcriptase (Promega) at 25 ºC for 10 min, 37 ºC for 60 min, 85 ºC for 5 min and 4 ºC for 4 min. The amplification was performed using the following primer sets: HTNV forward: 5'-TCTAGTT-GTATCCCCATCGACTG-3', HTNV reverse: 5'-ACATGC-GGAATACAATTATGGC-3'; human GAPDH forward: 5'-GGTGGTCCTCTGACTTCAACA-3', human GAPDH reverse: 5'-GTTGCTGTAGCCAAATTCGTTGT-3'; and mouse GAPDH forward: 5'-ACCCAGAAGACTGTGGA TGG-3', mouse GAPDH reverse: 5'-ACACATTGGGGGTAGGA-A CA-3'. The plasmid pGEM-T/HTNV, pGEM-T/human GAPDH, and pGEM-T/mouse GAPDH were stored in our laboratory and used to generate the standard curves. The absolute quantification of the HTNV viral gene was performed as previously described [20] . The data are presented as vRNA copies/ng of GAPDH mRNA.

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