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Whole Genome Amplification for Sequencing and Applications in Conservation Genetics
Author(s) -
JANEČKA JAN E.,
GRASSMAN LON I.,
HONEYCUTT RODNEY L.,
TEWES MICHAEL E.
Publication year - 2007
Publication title -
the journal of wildlife management
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.94
H-Index - 111
eISSN - 1937-2817
pISSN - 0022-541X
DOI - 10.2193/2006-369
Subject(s) - multiple displacement amplification , biology , amplicon , polymerase chain reaction , applications of pcr , genome , genomic dna , genetics , in silico pcr , dna sequencing , dna , dna nanoball sequencing , mitochondrial dna , microbiology and biotechnology , genomic library , digital polymerase chain reaction , multiplex polymerase chain reaction , gene , dna extraction , base sequence
We describe a method for rapidly amplifying whole genomes via a Phi29 DNA polymerase‐mediated strand displacement reaction (SDR). Genomic amplification products derived from the SDR reaction resulted in high quantities of DNA suitable for polymerase chain reaction (PCR) amplification and sequencing of mitochondrial genomes. Control region sequences of DNA derived directly from PCR amplicons of extracted DNA were identical to those derived from PCR amplification of SDR genomic DNA. Effective SDR amplification and subsequent sequencing was successful across tissues sources ranging in age from 1 year to 19 years. Strand replacement reaction genomic amplification offers a means of obtaining large quantities of DNA from small amounts of tissue.