Author: Soon Keong Wee; Suppiah Paramalingam Sivalingam; Eric Peng Huat Yap
Title: Rapid direct nucleic acid amplification test without RNA extraction for SARS-CoV-2 using a portable PCR thermocycler Document date: 2020_4_20
ID: e1oinu71_6
Snippet: Samples and controls used for assay development For biosafety purposes, synthetic nucleic acids rather than whole virus templates were used for the development of this assay. As SARS-CoV-2 is a positive stranded RNA coronavirus, we synthesized single stranded RNA (ssRNA) of the amplicon sequence (99 bp) for SARS-CoV-2 nucleocapsid (N) gene (Integrated DNA Technologies, San Diego). Plasmid DNA containing the N gene of SARS-CoV-2, MERS-CoV and SARS.....
Document: Samples and controls used for assay development For biosafety purposes, synthetic nucleic acids rather than whole virus templates were used for the development of this assay. As SARS-CoV-2 is a positive stranded RNA coronavirus, we synthesized single stranded RNA (ssRNA) of the amplicon sequence (99 bp) for SARS-CoV-2 nucleocapsid (N) gene (Integrated DNA Technologies, San Diego). Plasmid DNA containing the N gene of SARS-CoV-2, MERS-CoV and SARS-CoV were also used as positive controls (2019-nCoV RUO Plasmid Controls, Integrated DNA Technologies, San Diego). ssRNA of human ribonuclease P (RP) gene amplicon sequence (65 bp) was synthesized for use as internal control (Integrated DNA Technologies, San Diego).
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