RahmanJaquishKhasa2000

Référence

Rahman, M.H., Jaquish, B., Khasa, D.P. (2000) Optimization of PCR Protocol in Microsatellite Analysis with Silver and SYBR. Plant Molecular Biology Reporter, 18:339-348.

Résumé

Here we present the optimization of PCR conditions for microsatellite analysis of coniferous trees. The use of touchdown protocol for annealing resulted in a high success rate for optimization using fewer temperature profiles. The use of SYBR® Green gel stain to detect PCR products in agarose gels was more sensitive than ethidium bromide. This is valuable for determining the success of PCR reactions and estimating the amount of PCR products formed — which is crucial in determining the dilution required to produce bands of similar intensity upon silver staining of the polyacrylamide gels. The use of SYBR® Gold for staining polyacrylamide gels was not satisfactory in terms of the image quality produced. However, it was comparable to silver staining in terms of sensitivity, and could possibly be used in cases where the products are present as sharp single bands. In those cases, the use of SYBR® Gold gel stain would save time and money for staining polyacrylamide gels.

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@ARTICLE { RahmanJaquishKhasa2000,
    AUTHOR = { Rahman, M.H. and Jaquish, B. and Khasa, D.P. },
    TITLE = { Optimization of PCR Protocol in Microsatellite Analysis with Silver and SYBR },
    JOURNAL = { Plant Molecular Biology Reporter },
    YEAR = { 2000 },
    VOLUME = { 18 },
    PAGES = { 339-348 },
    ABSTRACT = { Here we present the optimization of PCR conditions for microsatellite analysis of coniferous trees. The use of touchdown protocol for annealing resulted in a high success rate for optimization using fewer temperature profiles. The use of SYBR® Green gel stain to detect PCR products in agarose gels was more sensitive than ethidium bromide. This is valuable for determining the success of PCR reactions and estimating the amount of PCR products formed — which is crucial in determining the dilution required to produce bands of similar intensity upon silver staining of the polyacrylamide gels. The use of SYBR® Gold for staining polyacrylamide gels was not satisfactory in terms of the image quality produced. However, it was comparable to silver staining in terms of sensitivity, and could possibly be used in cases where the products are present as sharp single bands. In those cases, the use of SYBR® Gold gel stain would save time and money for staining polyacrylamide gels. },
    KEYWORDS = { microsatellite, optimization, PCR, silver stain, SYBR® stain },
    OWNER = { brugerolles },
    TIMESTAMP = { 2007.12.05 },
}

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