Simultaneous amplification of 5′ and 3′ cDNA ends based on template-switching effect and inverse PCR | Zendy

Junchao Huang | Zendy; Feng Chen | Zendy

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Simultaneous amplification of 5′ and 3′ cDNA ends based on template-switching effect and inverse PCR

Author(s) -

Junchao Huang,

Feng Chen

Publication year - 2006

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/000112051

Subject(s) - complementary dna , inverse polymerase chain reaction , microbiology and biotechnology , biology , polymerase chain reaction , inverse , genetics , computational biology , gene , nested polymerase chain reaction , mathematics , geometry

A new approach for simultaneously amplifying the 5' and 3' ends of a desired cDNA is described. The method combines the template-switching effect with inverse PCR, which generates the flanking 5' and 3' regions of a certain cDNA with very low or no background. It requires only minimal amounts of total RNA for the synthesis of first-strand cDNA, while the same cDNA can be used to amplify flanking sequences of any cDNA species present in the sample. This method is reliable and easy to perform, which is very useful for isolating cDNA species of rare transcripts.

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