Author: Zhou, Yan; Zheng, HaiHong; Gao, Fei; Tian, DeBin; Yuan, ShiShan
Title: Mutational analysis of the SDD sequence motif of a PRRSV RNA-dependent RNA polymerase Document date: 2011_9_16
ID: yo9libo0_15
Snippet: Total intracellular RNAs were isolated using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. RNAs were suspended in RNase-free water, aliquoted, and stored at ï€70°C. For the analysis of virus transcription, the subgenomic (sg) mRNA7s of PRRSV was detected by reverse transcription-PCR (RT-PCR) from the total intracellular RNAs of transfected cells ( Figure 2 ). As an internal control for RT-PCR analy.....
Document: Total intracellular RNAs were isolated using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. RNAs were suspended in RNase-free water, aliquoted, and stored at ï€70°C. For the analysis of virus transcription, the subgenomic (sg) mRNA7s of PRRSV was detected by reverse transcription-PCR (RT-PCR) from the total intracellular RNAs of transfected cells ( Figure 2 ). As an internal control for RT-PCR analysis of sg mRNA7s, the β-actin gene of BHK-21 cells was also detected using a pair of primers, actin-F and actin-R. For the sg mRNA7s RT reaction, primer SR15284 was used. For the sg mRNA7s PCR, primers SF12 and SR15284 were used. The PCR conditions comprised 18 cycles of 95°C denaturation (30 s), 63°C annealing (30 s), and 72°C extension (30 s), followed by incubation at 72°C for 10 min. The 18 cycles were followed by a 5-min incubation at 95°C.
Search related documents:
Co phrase search for related documents- β actin gene and RT PCR analysis: 1, 2
- internal control and rnase free water: 1, 2, 3
- internal control and RT PCR analysis: 1, 2, 3, 4
- intracellular rna and total intracellular rna: 1, 2, 3, 4, 5, 6
- reverse transcription RT PCR PCR detect and RT PCR analysis: 1
Co phrase search for related documents, hyperlinks ordered by date