An Improved Method of Reverse Transcription and Polymerase Chain Reaction (RT-PCR) to Efficiently Detect Potyvirus, Sweet potato feathery mottle virus (SPFMV) RNA in Sweet Potato (Short Communication)

概要

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To improve the detection method for SPFMV in sweet potato leaves, we compared three RNA extraction methods that utilized either guanidine thiocyanate-phenol (ISOGEN^<TM>), SDS-phenol or a CF-11 cellulose column. The first method gave the highest yield of total RNA (39.0 μg from 0.1 g leaf sample) with the largest potential to synthesize cDNA by reverse transcription with the shortest extraction time. We then compared three combinations of polymerases for RT-PCR. AMV reverse transcriptase followed by PCR with rTaq DNA polymerase gave the most sensitive detection, in which the amplified DNA band was detected in a sample containing 0.2 pg of total RNA. (Received August 4, 2000 ; Accepted December 7, 2000)
日本植物病理学会の論文
2001-05-25