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Single-strand conformational polymorphism (SSCP) analysis is a simple and sensitive technique for
mutation detection and genotyping. The principle of SSCP analysis is based on the fact that single-
stranded DNA has a defined conformation. Altered conformation due to a single base change in the
sequence can cause single-stranded DNA to migrate differently under nondenaturing electrophoresis
conditions. Therefore wild-type and mutant DNA samples display different band patterns.
(4) detection of mobility difference of the single-stranded DNAs by electrophoresis under non-
denaturing conditions.
Several methods have been developed to visualize the SSCP mobility shifts. These include
The use of SSCP analysis to discover and genotype single-nucleotide polymorphisms (SNPs) has
been widely applied to the genetics of hypertension, including both monogenic (e.g., Liddle's
syndrome) and polygenic disorders (e.g., essential hypertension).
What is SSCP?
SSCP Analysis: Single-Strand Conformation Polymorphism Analysis
SSCP is the simplest and most used method of mutation detection. PCR is
used to amplify the region of interest and the resultant DNA is separated as
single-stranded molecules by electrophoresis in a non-denaturing
polyacrylamide gel (Orita et al,1989). A strand of single-stranded DNA folds
differently from another if it differs by a single base, and it is believed that
mutation-induced changes of tertiary structure of the DNA results in
different mobilities for the two strands. These mutations are detected as the
appearance of new bands on autoradiograms (radioactive detection), by
silver staining of bands or the use of fluorescent PCR primers which are
subsequently detected by an automated DNA sequencer (non-radioactive
detection).
Advantages
Limitations
SSCP screening only tells you that a mutation exists. You must perform
subsequent DNA sequencing to determine the nature of the mutation that
caused an electrophoretic mobility shift in a given sample.