Rapid determination of the complexity of cDNA bands extracted from DDRT- PCR polyacrylamide gels | Zendy

Neil R. Smith | Zendy

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Rapid determination of the complexity of cDNA bands extracted from DDRT- PCR polyacrylamide gels

Author(s) -

Neil R. Smith

Publication year - 1997

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/25.17.3552

Subject(s) - complementary dna , biology , polyacrylamide , restriction enzyme , polyacrylamide gel electrophoresis , fingerprint (computing) , microbiology and biotechnology , gel electrophoresis , differential display , dna , genetics , enzyme , biochemistry , gene , artificial intelligence , computer science

A band extracted from a differential display polyacrylamide gel often represents a composite of heterogeneous products. We have developed a non- radioactive method to simply and rapidly analyse its complexity. A fluorescent restriction enzyme fingerprint of the composite mixture is generated. The number of individual bands observed in this fingerprint indicates the complexity of the re-amplified cDNA mixture. Restriction fingerprints of the inserts of cDNA subclones derived from the re-amplified cDNA mixture are compared to the composite fingerprint to select those representing the most intense bands in the composite. This dramatically reduces the number of clones required for further characterisation.

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