Isolation of genomic sequences flanking a retroviral insertion site using a novel PCR-based method.
A novel PCR approach for the isolation of genomic sequences that flank retroviral insertion sites uses two standard primers and defined PCR conditions. The method is based on the directional cloning of digested genomic DNA into the multiple cloning site of pUC-based plasmids, isolation of recombinant plasmid DNA from pooled colonies and PCR amplification using primers specific for the pUC vector and retroviral LTR sequences. The method is simple and can be adapted to enable rapid isolation of any genomic sequences tagged by insertion of foreign DNA.
Natarajan
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Department of Genetics, University of Cambridge, UK.
Boulter
C A
CA
D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D016678:Genome; D051379:Mice; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction / Q000379:methods*; D012190:Retroviridae / Q000235:genetics*
D017931:DNA Primers