Selected article for: "Extraction kit and Gel Extraction kit"
Author: Zhang, Fanfan; Ye, Yu; Song, Deping; Guo, Nannan; Peng, Qi; Li, Anqi; Zhou, Xingrong; Chen, Yanjun; Zhang, Min; Huang, Dongyan; Tang, Yuxin
Title: A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification
  • Document date: 2017_9_21
    • ID: 1k9l0z5k_25
    Snippet: In order to construct the standards of PDCoV N gene, the targeted RNA of PDCoV was reversely transcribed into single-stranded cDNA by a random primer and then amplified by PCR using forward primer PDCoV-NWF and reverse primer PDCoV-NWR ( Table 2 ). The reaction conditions were as follows: denaturation at 95 °C for 5 min, 38 cycles of 94 °C for 30 s, 53 °C for 30 s, 72 °C for 2 min, and a final extension at 72 °C for 7 min. The amplified prod.....
    Document: In order to construct the standards of PDCoV N gene, the targeted RNA of PDCoV was reversely transcribed into single-stranded cDNA by a random primer and then amplified by PCR using forward primer PDCoV-NWF and reverse primer PDCoV-NWR ( Table 2 ). The reaction conditions were as follows: denaturation at 95 °C for 5 min, 38 cycles of 94 °C for 30 s, 53 °C for 30 s, 72 °C for 2 min, and a final extension at 72 °C for 7 min. The amplified products were purified by E.Z.N.A ™ Gel Extraction Kit (Omega, USA) and subsequently cloned into E. coli JM109 using the pGEM-T easy vector (Promega, USA). The recombinant plasmid was extracted using TIANprep Mini Plasmid Kit (TIANGEN, China) following the protocol of the manufacturer. The concentration and quality of the plasmid DNA was determined by NanoDrop 2000 spectrophotometer (Thermo scientific, USA), which was then used as the standards for the quantitative analysis.

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