While conventional SSCP and heteroduplex analysis use
polyacrylamide gel electrophoresis to separate PCR products,
DHPLC uses a high pressure system to force the products
through a column under partially denaturing conditions.
Conditions for optimum separation of normal and mutant
sequences are created by the use of buffer gradients and
specific temperatures. The DNA molecules that are
progressively eluted from the column are monitored by an
ultraviolet detector with data being collected by computer.
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