2002
DOI: 10.1016/s0166-0934(01)00391-3
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Sodium sulphite inhibition of potato and cherry polyphenolics in nucleic acid extraction for virus detection by RT-PCR

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Cited by 72 publications
(41 citation statements)
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“…This problem could be avoided using commercial kits for purification of nucleic acids, but their cost is prohibitive for detecting the virus in epidemiological studies due to the large number of samples that need to be processed. These inhibitors are very common in plants, but do not affect molecular hybridization procedures (Singh et al, 2002). Tissue-print hybridization was consistent and permitted detection of PMoV-T on imprints from all infected tomato plants collected from the appropriate selected material, and turned out to be as robust as dot-blot hybridization.…”
Section: Discussionmentioning
confidence: 82%
“…This problem could be avoided using commercial kits for purification of nucleic acids, but their cost is prohibitive for detecting the virus in epidemiological studies due to the large number of samples that need to be processed. These inhibitors are very common in plants, but do not affect molecular hybridization procedures (Singh et al, 2002). Tissue-print hybridization was consistent and permitted detection of PMoV-T on imprints from all infected tomato plants collected from the appropriate selected material, and turned out to be as robust as dot-blot hybridization.…”
Section: Discussionmentioning
confidence: 82%
“…Due to the advantages of RT-PCR system (single/multiplex) concerning on time, cost, reliability and sensitivity, it has been documented as an alternative system for certification of plant propagating materials. Disadvantages of this system including the presence of inhibitors such as polyphenolics, polysaccharides and endogenous ribonucleases in the plant extracts (20) and inefficient synthesis of viroid genome (3) could be easily removed by proper RNA extraction methods and using proper sequence primers. Moreover, the accuracy and reliability of the extracted RNA and subsequently cDNA synthesis could be analyzed by the coamplification of the internal control (Nad 5 gene) in both d-and m-RT PCR.…”
Section: Discussionmentioning
confidence: 99%
“…Samples were stored at 4°C. Various other DNA extraction protocols were used on plant samples to remove inhibitors [20][21][22][23][24], including the PowerPlant Pro DNA isolation Kit (MO BIO Laboratories Inc., Carlsbad, CA).…”
Section: Dna Extractionmentioning
confidence: 99%