Reverse Dot Blot (RDB)
Allele-specific oligonucleotide (ASO) probes, complementary to mutant and normal DNA sequences, are bound to membranes (dots or slots). DNA that has been amplified in multiplex reaction(s) with biotinylated primers is hybridized to the immobilized oligonucleotides on the membrane. Hybridisation is detected by adding streptavidin-horseradish peroxidase.
View an animation of the Reverse Dot Blot.
Advantages
Detection is non-radioactive, convenient, rapid, robust and requires no specialized interpretation skills.
Disadvantages
Relatively inflexible and not easily expanded to include additional mutations.
Example of a commercial kit
INNO-LiPA CFTR19, INNO-LiPA CFTR17+Tn Update (Innogenetics
)
Last changed: 2010-01-25
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