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. 1994 Mar 1;91(5):1946–1949. doi: 10.1073/pnas.91.5.1946

Sequence-specific "gene signatures" can be obtained by PCR with single specific primers at low stringency.

S D Pena 1, G Barreto 1, A R Vago 1, L De Marco 1, F C Reinach 1, E Dias Neto 1, A J Simpson 1
PMCID: PMC43281  PMID: 8127912

Abstract

Low-stringency single specific primer PCR (LSSP-PCR) is an extremely simple PCR-based technique that detects single or multiple mutations in gene-sized DNA fragments. A purified DNA fragment is subjected to PCR using high concentrations of a single specific oligonucleotide primer, large amounts of Taq polymerase, and a very low annealing temperature. Under these conditions the primer hybridizes specifically to its complementary region and nonspecifically to multiple sites within the fragment, in a sequence-dependent manner, producing a heterogeneous set of reaction products resolvable by electrophoresis. The complex banding pattern obtained is significantly altered by even a single-base change and thus constitutes a unique "gene signature." Therefore LSSP-PCR will have almost unlimited application in all fields of genetics and molecular medicine where rapid and sensitive detection of mutations and sequence variations is important. The usefulness of LSSP-PCR is illustrated by applications in the study of mutants of smooth muscle myosin light chain, analysis of a family with X-linked nephrogenic diabetes insipidus, and identity testing using human mitochondrial DNA.

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Selected References

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