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Construction of Infectious Clones for RNA Viruses: TMV

 Last Update: 2020-11-21
 Source: Internet
 Author: User

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The generation of infectious clones is routinely the first step for reverse genetic studies of RNA plant virus gene and sequence function. The procedure given here, details the creation of c
DNA
clones of tobacco mosaic virus, from which infectious transcripts can be generated in vitro with T7 RNA polymerase. The procedure describes methods for virion purification, viral RNA extraction, reverse transcription,
PCR
amplification of genomic
cDNA
fragments, generation of a full-length cDNA clone under the control of a T7 promoter, in vitro transcription, and infectivity testing.

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