Amplification and cloning of a long RNA virus genome using immunocapture-long RT-PCR
A rapid and easy method was developed in order to amplify long fragments of the genome of potato virus Y (PVY), a virus possesing a 10-kb genomic RNA. The method of immunocapture-RT-PCR was adapted, by using thermostable DNA polymerases with proofreading activity and the proper buffers and cycles, to amplify almost the whole genome of PVY in two fragments (5.6 and 4.3 kb) without purifying virions nor viral RNA. Both fragments were cloned subsequently and their ends sequenced. The method is applicable to the rapid cloning and molecular characterization of the genomes of many other RNA viruses.
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| Main Authors: | , , |
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| Format: | journal article biblioteca |
| Language: | English |
| Published: |
Elsevier
1997
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| Subjects: | Immunocapture, Long RT-PCR, PVY, |
| Online Access: | http://hdl.handle.net/20.500.12792/3880 http://hdl.handle.net/10261/294463 |
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