Reverse Dot Blot

Reverse dot blots entail a hybridization-based technology in which the oligonucleotide probes complementary to the normal and mutant sequences are attached to a membrane in a miniarray format. Patient DNA is subjected to one or more multiplex PCR reactions using biotin-tagged primers for all exons containing mutations of interest, and the resulting biotinylated PCR products (the target) are hybridized to the array (the probes). Both mutant and wildtype alleles are interrogated simultaneously. A streptavi-din-peroxidase conjugate complex, which binds to the biotinylated PCR product hybridized to the arrayed oli-gonucleotide probe, is used to visualize the test results.[12] Current commercially available versions of this technology (Roche, Innogenetics) incorporate the 25 mutations and variants (5T/7T/9T) recommended for population screening. The reverse blot platform has numerous advantages including direct genotyping, nonisotopic detection, and the potential for automation.

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

Get My Free Ebook


Post a comment