Selected article for: "cdna synthesis and extraction rna extraction"
Author: Zhou, Jie; Li, Cun; Sachs, Norman; Chiu, Man Chun; Wong, Bosco Ho-Yin; Chu, Hin; Poon, Vincent Kwok-Man; Wang, Dong; Zhao, Xiaoyu; Wen, Lei; Song, Wenjun; Yuan, Shuofeng; Wong, Kenneth Kak-Yuen; Chan, Jasper Fuk-Woo; To, Kelvin Kai-Wang; Chen, Honglin; Clevers, Hans; Yuen, Kwok-Yung
Title: Differentiated human airway organoids to assess infectivity of emerging influenza virus
  • Document date: 2018_6_26
    • ID: z637eh2z_23
    Snippet: RNA Extraction and RT-qPCR. To evaluate the differentiation status of AOs cultured in PD medium versus those in AO medium, the organoids were harvested at the indicated times, and RNA was extracted using MiniBEST Universal RNA Extraction Kit (TaKaRa). To quantify virus replication, the organoids and supernatant samples were lysed for RNA extraction using the MiniBEST Universal RNA Extraction Kit and MiniBEST Viral RNA/DNA Extraction Kit (TaKaRa),.....
    Document: RNA Extraction and RT-qPCR. To evaluate the differentiation status of AOs cultured in PD medium versus those in AO medium, the organoids were harvested at the indicated times, and RNA was extracted using MiniBEST Universal RNA Extraction Kit (TaKaRa). To quantify virus replication, the organoids and supernatant samples were lysed for RNA extraction using the MiniBEST Universal RNA Extraction Kit and MiniBEST Viral RNA/DNA Extraction Kit (TaKaRa), respectively. cDNA was synthesized with the Transcriptor First Strand cDNA Synthesis Kit (Roche) with Oligo-dT primer. qPCR was performed with LightCycler 480 SYBR Green I Master Mix (Roche) using genespecific primers (SI Appendix, Table S2 ) to detect cellular gene-expression level and viral gene copy number. The mRNA expression levels of cellular genes were normalized with that of GAPDH. Viral gene copy number was determined by absolute quantification using a plasmid expressing a conserved region of the IAV M gene.

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