Technique / Genetics / Genetic Screening / SSCP Single-strand conformation polymorphism

Single strand conformation polymorphism (SSCP) is defined as conformational difference of single stranded nucleotide sequences of identical length as induced by differences in the sequences under certain experimental conditions. This property allows to distinguish the sequences by means of gel electrophoresis, which separates the different conformations.

Single-strand conformation polymorphism analysis using capillary electrophoresis.
(Curr Protoc Hum Genet. 2003 May;Chapter 7:Unit 7.12.)
Larsen LA, Christiansen M, Vuust J, Andersen PS.

Statens Serum Institut, Copenhagen, Denmark.

Single-strand conformation polymorphism (SSCP) is one of the most frequently used
mutation detection methods. This unit describes a method of SSCP with automated
analysis by capillary electrophoresis in order to increase the capacity and
throughput. A protocol is provided for sample preparation. For a medium
throughput laboratory, a single capillary instrument, as described in this unit,
may be quite sufficient. In many cases, however, screening for mutations in large
population groups requires a high throughput, and this is best obtained through
the use of a multi-capillary instrument, as discussed.

Detection of mutations by single-strand conformation polymorphism (SSCP) analysis
(Curr Protoc Hum Genet. 2001 May;Chapter 7:Unit 7.4.)
Warren W, Hovig E, Smith-Sørensen B, Børresen AL, Fujimura FK, Liu Q, Feng J,
Sommer SS.

Institute of Cancer Research, Surrey, UK.

Single-strand conformation polymorphism (SSCP) analysis detects mutations based
on the fact that single-nucleotide changes in DNA sequences alter the mobility of
single-stranded DNA in nondenaturing gels. Four methods for detecting mutations
based on SSCP are described here. (1) Traditional SSCP analysis is technically
easy and can be used for screening large numbers of samples. SSCP-hybrid methods
detect mutations based on either an SSCP effect or an altered component
independent of the SSCP effect. (2) Dideoxy fingerprinting (ddF) involves PCR
amplification of the target and creation of a set of dideoxy-terminated strands
with the mutation. (3) Bi-directional dideoxy fingerprinting (Bi-ddF) involves
production of two sets of dideoxy-terminated strands that are generated from two
different primers. (4) Restriction endonuclease fingerprinting (REF) involves
cleavage of the amplified target with five to six groups of restriction
endonucleases.