Author: Azeem Mehmood Butt; Shafiqa Siddique; Xiaoping An; Yigang Tong
Title: Development of a dual-gene loop-mediated isothermal amplification (LAMP) detection assay for SARS-CoV-2: A preliminary study Document date: 2020_4_11
ID: fc338qdt_15
Snippet: The eluted viral RNA was reverse transcribed by using Hifair II 1 st stand cDNA Supermix (Yeasen, China) as per manufacturer's recommendations. Briefly, 10ul of Supermix containing optimized concentration of Hifair II reverse transcriptase, RNase inhibitor, dNTPs, random hexamer/oligo dT primer mix in an optimized buffer system was mixed with 5ul of Viral RNA followed by addition of 5ul DNase/RNase free water (Invitrogen, USA) for a final reactio.....
Document: The eluted viral RNA was reverse transcribed by using Hifair II 1 st stand cDNA Supermix (Yeasen, China) as per manufacturer's recommendations. Briefly, 10ul of Supermix containing optimized concentration of Hifair II reverse transcriptase, RNase inhibitor, dNTPs, random hexamer/oligo dT primer mix in an optimized buffer system was mixed with 5ul of Viral RNA followed by addition of 5ul DNase/RNase free water (Invitrogen, USA) for a final reaction volume of 20ul. The procedure was performed on MiniAmp thermocycler (Applied biosystems, USA) and the cycling conditions were 25°C for 5min followed by 42°C for 30min and final elongation/termination at 85°C for 5min.
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