We describe a single step reverse transcription polymerase chain reaction protocol that can be used to amplify part of the neuraminidase gene segment (segment 6) from all nine subtypes of influenza A virus. The method has also been applied to amplify gene segment 1 of influenza A, which encodes the basic polymerase protein 2 (PB2). The method combines the use of mixed base primers with a "touchdown" thermal cycling program and is applicable to a wide range of nucleic acid targets in which there is genetic variability in the regions complementary to the PCR primers.
CITATION STYLE
Alvarez, A. C., Boyd, V., Lai, R., Pineda, S., Bletchly, C., Heine, H. G., & Barnard, R. (2010). Detection of influenza A virus neuraminidase and PB2 gene segments by one step reverse transcription polymerase chain reaction. Methods in Molecular Biology (Clifton, N.J.), 630, 65–81. https://doi.org/10.1007/978-1-60761-629-0_5
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