Sequencing By Hybridization refers to a technique used in molecular biology and genetics to determine the sequence of a specific DNA or RNA molecule. It involves utilizing the principle of complementary base pairing between DNA or RNA strands to identify the sequence information.
In this method, a mixture of different short oligonucleotide probes, usually around 20 to 30 nucleotides in length, are designed to be complementary to specific regions of the target DNA or RNA molecule. These probes are labeled with different fluorescent dyes or other detectable markers for identification.
The first step in the Sequencing By Hybridization technique involves denaturing the target DNA or RNA into single-stranded molecules. Then, the labeled oligonucleotide probes are mixed with the sample and allowed to hybridize or bind to their complementary sequences.
Once hybridization has occurred, the sample is washed to remove any unbound probes. The labeled probes that have successfully hybridized to their complementary sequences can be detected using fluorescence or any other applicable detection method.
By comparing the sequence of the labeled probes that have bound to the target DNA or RNA molecule, the original sequence of the molecule can be inferred. The analysis of the signal intensity of each probe can provide information about the order of nucleotides in the target sequence.
Overall, Sequencing By Hybridization is a powerful tool that allows for the determination of DNA or RNA sequences by exploiting the principles of complementary base pairing. It can be used for a wide range of applications, including genome mapping, mutation detection, and gene expression analysis.
