Selected article for: "agarose gel and gel cut"
Author: Gun-Soo Park; Keunbon Ku; Seung-Hwa Beak; Seong Jun Kim; Seung Il Kim; Bum-Tae Kim; Jin-Soo Maeng
Title: Development of Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) Assays Targeting SARS-CoV-2
  • Document date: 2020_3_12
    • ID: 1qn5y4gc_12
    Snippet: To prepare standard RNA for hCoV-229E, first amplicon of PCR (Forward primer: 5' -GCTAGTGGATGATCATGCTTTG -3', Reverse primer: 5' -TGGGGCCATAAACTGTTCTATTAC -3') was cloned to pBluescript II KS (+) plasmid with BamHI and XhoI. Then, in vitro transcription template was prepared by restriction enzyme cut with BglI and XhoI and purification after agarose gel electrophoresis. For hCoV-OC43 and MERS-CoV, amplicons of PCR (OC43 forward primer: 5' -AGCAAC.....
    Document: To prepare standard RNA for hCoV-229E, first amplicon of PCR (Forward primer: 5' -GCTAGTGGATGATCATGCTTTG -3', Reverse primer: 5' -TGGGGCCATAAACTGTTCTATTAC -3') was cloned to pBluescript II KS (+) plasmid with BamHI and XhoI. Then, in vitro transcription template was prepared by restriction enzyme cut with BglI and XhoI and purification after agarose gel electrophoresis. For hCoV-OC43 and MERS-CoV, amplicons of PCR (OC43 forward primer: 5' -AGCAACCAGGCTGATGTCAATACC -3', OC43 reverse primer: 5' -AGCAGACCTTCCTGAGCCTTCAAT -3', MERS-CoV: UpE region [21] ) were synthesized and cloned to pBIC-A plasmid (Bioneer). In vitro transcription template for hCoV-OC43 and MERS-CoV were prepared by restriction enzyme cut with BamHI-XhoI or SspI-XhoI, respectively. In vitro transcriptions were done with EZâ„¢ T7 High Yield In Vitro Transcription kit (Enzynomics) as manufacturer's instructions. RNA products were then purified using Agencourt RNAClean XP (Beckman Coulter).

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