PCR-generated padlock probes detect single nucleotide variation in genomic DNA
2000 (English)In: Nucleic Acids Research, ISSN 0305-1048, Vol. 28, no 12, E58- p.Article in journal (Refereed) Published
Circularizing oligonucleotide probes, so-called padlock probes, have properties that should prove valuable in a wide range of genetic investigations, including in situ analyses, genotyping and measurement of gene expression. However, padlock probes can be difficult to obtain by standard oligonucleotide synthesis because they are relatively long and require intact 5'- and 3'-end sequences to function. We describe a PCR-based protocol for flexible small-scale enzymatic synthesis of such probes. The protocol also offers the advantage over chemical synthesis that longer probes can be made that are densely labeled with detectable functions, resulting in an increased detection signal. The utility of probes synthesized according to this protocol is demonstrated for the analysis of single nucleotide variations in human genomic DNA both in situ and in solution.
Place, publisher, year, edition, pages
2000. Vol. 28, no 12, E58- p.
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-51440PubMedID: 10871381 OAI: oai:DiVA.org:uu-51440DiVA: diva2:79349