Barnes' protocol for long PCR to incorporate a high level of an exonuclease-free, N-terminal deletion mutant of Taq DNA polymerase in combination with a very low level of a thermostable DNA polymerase exhibiting a 3' exonuclease activity. The reaction was modified to include a reverse-transcription step and was optimized for use with hepatitis C virus genomes as template. It has been possible to amplify large amounts of 4 to 4.8 kb of hepatitis C virus genome from clinical samples. This amplified cDNA will be sequenced to complete genotyping studies of the virus, and used for the construction of full-length, possibly infectious, cDNA molecules.
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| Puskas, John; Skrombolas, Denise; Sedlacek, Abigail et al. (2011) Development of an attenuated interleukin-2 fusion protein that can be activated by tumour-expressed proteases. Immunology 133:206-20 |
| Taylor, Simon R J; Gonzalez-Begne, Mireya; Sojka, Dorothy K et al. (2009) Lymphocytes from P2X7-deficient mice exhibit enhanced P2X7 responses. J Leukoc Biol 85:978-86 |
