Author: Si, Wei; Zhou, Shun; Wang, Zhao; Cui, Shang-jin
Title: A multiplex reverse transcription-nested polymerase chain reaction for detection and differentiation of wild-type and vaccine strains of canine distemper virus Document date: 2010_5_1
ID: 1fw0xiin_5
Snippet: Total RNA was extracted from the infected cells with TRIzol ® reagent in accordance with the manufacturer's instructions. cDNA synthesis reaction was performed by polymerase chain reaction as described elsewhere using primer Moloney murine leukemia virus reverse transcriptase (M-MLV RT) [6] [7] [8] [9] [10] . The first-round PCR was performed with primers P1 and P4. A nested PCR was performed in a total volume of 25 μL containing the firstround.....
Document: Total RNA was extracted from the infected cells with TRIzol ® reagent in accordance with the manufacturer's instructions. cDNA synthesis reaction was performed by polymerase chain reaction as described elsewhere using primer Moloney murine leukemia virus reverse transcriptase (M-MLV RT) [6] [7] [8] [9] [10] . The first-round PCR was performed with primers P1 and P4. A nested PCR was performed in a total volume of 25 μL containing the firstround PCR products diluted tenfold as well as each of primers P2/P4, P3/P4, or P2, P3/P4 [10] [11] [12] . The RT-nPCR products were visualized by electrophoresis in a 2% (w/v) agarose gel.
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