Studies on cDNA Probe Detection Technology of Apple stem pitting virus in Korla Pear

摘要

Using leaves and phloem of Korla pear as materials, we extracted total RNA through two methods. By comparison, superior isolation method of extracting total RNA form Korla pear is screened out, and the cDNA Probe labeled by Biotin is yielded with RT-PCR. The highest sensitivity of hybridization is obtained at probe concentration 400ng/mL, 45% formamide and 42℃ for 6 hours. The best result of hybridization is import nitrocellulose membrane. Blocking result of Tween20 is better than that of Album in Bovine. This study indicates that the detection of the total RNA extracted by two methods from fresh leaf, old leaf, phloem and freezing leaf shows hybridization signal.