Anchor primer associated problems in differential display reverse transcription polymerase chain reaction

Article


Chen, Yihua, Wang, Bin, Weining, Song and Daggard, Grant. 2004. "Anchor primer associated problems in differential display reverse transcription polymerase chain reaction." Analytical Biochemistry. 329 (1), pp. 145-147. https://doi.org/10.1016/j.ab.2004.02.030
Article Title

Anchor primer associated problems in differential display reverse transcription polymerase chain reaction

ERA Journal ID1611
Article CategoryArticle
AuthorsChen, Yihua (Author), Wang, Bin (Author), Weining, Song (Author) and Daggard, Grant (Author)
Journal TitleAnalytical Biochemistry
Journal Citation329 (1), pp. 145-147
Number of Pages3
Year2004
Place of PublicationUnited States
ISSN0003-2697
1096-0309
Digital Object Identifier (DOI)https://doi.org/10.1016/j.ab.2004.02.030
Web Address (URL)https://www.sciencedirect.com/science/article/pii/S0003269704002064
Abstract

Differential display reverse transcription PCR (DDRT-PCR)2 was first reported by P. Liang and A.B. Paredee in 1992. The method utilized an anchor primer, including a poly(T) component with the addition of one or two select bases in reverse transcription. The first-strand cDNA was amplified via PCR using the same anchor primer and a 10-base random primer. As the DDRT-PCR method is simple and sensitive, it has become a popular technology in gene expression work. However, a major problem with the technique is its ability
to produce a high rate of false-positive products. A number of approaches to overcome such problems have been suggested, including using improved gel resolution systems, performing PCR with the cDNA generated by two different reverse transcriptases, and using cytoplasmic RNA to avoid unprocessed mRNA. While there has been a focus in the literature on modification of the random primer component as a means of improving DDRT-PCR reliability, in this study we report on the impact of modifications to the traditional anchor primer component and the role of anchor primers. The addition of weight bases is important for improving the reliability of traditional anchor primer in the DDRT PCR condition and can contribute to the reproducibility of DDRT PCR products.

KeywordsDDRT-PCR, differential display reverse transcriptionPCR
Contains Sensitive ContentDoes not contain sensitive content
ANZSRC Field of Research 2020300199. Agricultural biotechnology not elsewhere classified
310505. Gene expression (incl. microarray and other genome-wide approaches)
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Byline AffiliationsFaculty of Sciences
Southwest University of Science and Technology, China
Department of Primary Industries, Queensland
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