A simple method for direct cloning and sequencing cDNA by the use of a single specific oligonucleotide and oligo(dT) in a polymerase chain reaction (PCR)

cDNA cloning by PCR with the use of two mixed oligonucleotides corresponding to the same mRNA has been described recently. The method reported here was designed to clone the 3' region of the human parvalbumin gene but is suitable for cloning any cDNA especially when only one short amino acid (e...

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Bibliographic Details
Published in:Nucleic acids research 1989-01, Vol.17 (1), p.453-453
Main Author: BERCHTOLD, M. W
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biological and medical sciences
Cloning, Molecular - methods
Diverse techniques
DNA - genetics
Fundamental and applied biological sciences. Psychology
Genes
Molecular and cellular biology
nucleotide sequence
Oligodeoxyribonucleotides
Oligonucleotide Probes
Parvalbumins - genetics
polymerase chain reaction
Rats
Citations: Items that cite this one
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Description
Summary:cDNA cloning by PCR with the use of two mixed oligonucleotides corresponding to the same mRNA has been described recently. The method reported here was designed to clone the 3' region of the human parvalbumin gene but is suitable for cloning any cDNA especially when only one short amino acid (eg. N-terminal of a protein) or nucleic acid sequence is known, when the abundance of the transcript of interest is low when the RNA source is limited. In addition, this method allows the amplification of sequences from cDNA libraries without the need for plating bacteria or phages.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/17.1.453