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Journal of Virological Methods
Bar-Joseph, M., Virus Laboratory, Volconi Center, A.R.O., Bet Dagan, Israel
Rosner, A., Virus Laboratory, Volconi Center, A.R.O., Bet Dagan, Israel
Moscovitz, M., Virus Laboratory, Volconi Center, A.R.O., Bet Dagan, Israel
Hull, R., Department of Virus Research, John Innes Institute, Colney Lane, Norwich, United Kingdom
A simple procedure for the isolation of double-stranded (ds) RNA from virus-infected plants is described. The method is based on grinding plant tissue in 4% p-aminosalicylic acid and recovery of ds RNA by phenol extraction and precipitation with 30% ethanol. The presence of both negative and positive virus RNA strands in RNA fractionated in agarose gels was verified by Northern blot hybridization with polynucleotide kinase labelled genomic RNA or complementary DNA (cDNA) probes. The procedure enabled detection of three major ds RNA species (MWs 4.2, 1.05 and 0.48 × 106) and at least 4 minor bands with estimated MWs of 3.5, 2.5, 2.2 and 2.0 × 106 in Nicotiana tabacum plants systemically infected with tobacco mosaic virus (TMV). Cucumber mosaic virus (CMV)-infected Pachystachys coccinea plants contained 2 minor bands of MWs 0.49 and 0.35 × 106 in addition to the previously described 4 major ds RNAs and ds CARNA 5 (MW 0.22 × 106). The patterns of ds RNA are useful for diagnosing natural infections of CMV and TMV in N. glauca plants and of citrus tristeza virus in Citrus spp. © 1983.
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תנאי שימוש
A simple procedure for the extraction of double-stranded RNA from virus-infected plants
6
Bar-Joseph, M., Virus Laboratory, Volconi Center, A.R.O., Bet Dagan, Israel
Rosner, A., Virus Laboratory, Volconi Center, A.R.O., Bet Dagan, Israel
Moscovitz, M., Virus Laboratory, Volconi Center, A.R.O., Bet Dagan, Israel
Hull, R., Department of Virus Research, John Innes Institute, Colney Lane, Norwich, United Kingdom
A simple procedure for the extraction of double-stranded RNA from virus-infected plants
A simple procedure for the isolation of double-stranded (ds) RNA from virus-infected plants is described. The method is based on grinding plant tissue in 4% p-aminosalicylic acid and recovery of ds RNA by phenol extraction and precipitation with 30% ethanol. The presence of both negative and positive virus RNA strands in RNA fractionated in agarose gels was verified by Northern blot hybridization with polynucleotide kinase labelled genomic RNA or complementary DNA (cDNA) probes. The procedure enabled detection of three major ds RNA species (MWs 4.2, 1.05 and 0.48 × 106) and at least 4 minor bands with estimated MWs of 3.5, 2.5, 2.2 and 2.0 × 106 in Nicotiana tabacum plants systemically infected with tobacco mosaic virus (TMV). Cucumber mosaic virus (CMV)-infected Pachystachys coccinea plants contained 2 minor bands of MWs 0.49 and 0.35 × 106 in addition to the previously described 4 major ds RNAs and ds CARNA 5 (MW 0.22 × 106). The patterns of ds RNA are useful for diagnosing natural infections of CMV and TMV in N. glauca plants and of citrus tristeza virus in Citrus spp. © 1983.
Scientific Publication
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