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Direct reverse transcription--polymerase chain reaction from whole blood without RNA extraction.

作者信息

Shi Y J, Liu J Z

机构信息

Department of Biochemistry and Molecular Biology, Chinese Academy of Medical Sciences, Beijing.

出版信息

Genet Anal Tech Appl. 1992 Oct-Dec;9(5-6):149-50. doi: 10.1016/1050-3862(92)90041-3.

DOI:10.1016/1050-3862(92)90041-3
PMID:1284274
Abstract

A simple, fast, efficient method of direct reverse transcription-polymerase chain reaction (RT-PCR) from whole blood without RNA extraction is described. After the erythrocytes in a few microliters of fresh blood were destroyed and removed, the total RNA was released from the leukocytes by means of water treated with diethyl pyrocarbonate. The solution was used as RNA template and directly introduced in the RT-PCR. Following this method, which is called direct RT-PCR, three different cDNA fragments were successfully amplified with three different pairs of primers, respectively. Meanwhile, the direct RT-PCR was compared with the RT-PCR from the extracted total RNA template. The results showed no obvious difference.

摘要

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