Details

Autor(en) / Beteiligte

• Grothues, Dietmar
• Cantor, Charles R.
• Smith, Cassandra L.

Titel

PCR amplification of megabase DNA with tagged random primers (T-PCR)

Ist Teil von

• Nucleic acids research, 1993-03, Vol.21 (5), p.1321-1322

Ort / Verlag

Oxford: Oxford University Press

Erscheinungsjahr

1993

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• A very sensitive and specific method for the random amplification of whole DNA molecules and genomes ranging from 400 base pairs (bp) to 40 Megabase (Mb) is described. This simple, two step PCR strategy utilizes tagged random primers that consist of a pool of all possible 3' sequences for binding to the target DNA and a constant 5' region for the detection of incorporated primers. As little as 10 super(-12) g of DNA was readily amplified to amounts that could be visualized on ethidium bromide stained gels. The high specificity of this method was demonstrated by hybridization of PCR products to gridded whole genome cosmid libraries. Tagged random primer PCR (T-PCR) is very useful for the amplification of DNA samples purified by various electrophoresis techniques or flow sorting. This random whole DNA and genome amplification should simplify the analysis of other samples with very little DNA, like individual sperm and oocytes. It has the potential for amplifying any DNA molecule.