Author: Nicola Clementi; Elena Criscuolo; Roberta Antonia Diotti; Roberto Ferrarese; Matteo Castelli; Roberto Burioni; Massimo Clementi; Nicasio Mancini
Title: Combined prophylactic and therapeutic use maximizes hydroxychloroquine anti-SARS-CoV-2 effects in vitro Document date: 2020_3_31
ID: fhy2z49t_12
Snippet: Viral RNA extraction and real-time RT-PCR. Viral RNA was purified from 140 mL of cell culture supernatant using the QIAamp Viral RNA Mini Kit (QIAGEN), following the manufacturer's instructions. Subsequently, the purified RNA was used to perform the synthesis of first-strand cDNA, using the SuperScript™ First-Strand Synthesis System for RT-PCR (Thermo Fisher Scientific), following the manufacturer's instruction. Real-time PCR, using SYBR® Gree.....
Document: Viral RNA extraction and real-time RT-PCR. Viral RNA was purified from 140 mL of cell culture supernatant using the QIAamp Viral RNA Mini Kit (QIAGEN), following the manufacturer's instructions. Subsequently, the purified RNA was used to perform the synthesis of first-strand cDNA, using the SuperScript™ First-Strand Synthesis System for RT-PCR (Thermo Fisher Scientific), following the manufacturer's instruction. Real-time PCR, using SYBR® Green dyebased PCR amplification and detection method, was performed in order to detect the cDNA. We used the SYBR™ Green PCR Master Mix (Thermo Fisher Scientific) the forward primer N2F: TTA CAA ACA TTG GCC GCA AA, the reverse primer N2R: GCG CGA CAT TCC GAA GAA, and the following PCR conditions: 95°C for 2 min, 45 cycles of 95°C for 20 sec, annealing at 55°C for 20 sec and elongation at 72°C for 30 sec, followed by a final elongation at 72°C for 10 min 7 . RT-PCR was performed using the ABI-PRISM 7900HT Fast Real-Time instruments (Applied Biosystems) by using optical-grade 96-well plates. Samples were run in duplicate in a total volume of 20 μL.
Search related documents:
Co phrase search for related documents- Applied Biosystems real time instrument and Fast Applied Biosystems real time instrument: 1, 2
- Applied Biosystems real time instrument and real time: 1, 2
- Applied Biosystems real time instrument and real time instrument: 1, 2
- Applied Biosystems real time instrument and real time pcr: 1, 2
- cdna detect and cell culture: 1
- cdna detect and PCR amplification: 1
- cdna detect and real time: 1, 2
- cdna detect and real time pcr: 1, 2
- cdna detect and Viral RNA extraction: 1
- cell culture and detection method: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20
- cell culture and Mini kit: 1, 2, 3, 4, 5, 6, 7
- cell culture and PCR amplification: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34
- cell culture and QIAamp viral RNA Mini kit: 1, 2, 3, 4
- cell culture and QIAGEN QIAamp viral RNA Mini kit: 1, 2, 3
- cell culture and real time: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74
- cell culture and real time instrument: 1
- cell culture and real time pcr: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74
- cell culture and strand cDNA: 1, 2, 3
- cell culture and strand cDNA synthesis: 1, 2
Co phrase search for related documents, hyperlinks ordered by date