Sequencing by Hybridizations, also known as SBH, is a method used in molecular biology for determining the sequence of nucleotides in a DNA or RNA molecule. It is specifically based on the principle of DNA hybridization, where the binding of complementary nucleotide sequences is exploited to infer the sequence.
The process of SBH involves several steps. First, a DNA or RNA molecule is fragmented into short pieces. These fragments are then labeled with a fluorescent tag or a radioisotope for detection. In a parallel step, a collection of DNA or RNA probes, each representing a specific nucleotide sequence, is prepared. These probes are also labeled with a fluorescent tag or a radioisotope.
Next, the labeled DNA or RNA fragments and the labeled probes are mixed together and allowed to hybridize. The binding of fragments to probes occurs when their sequences are complementary, forming a double-stranded helix. By employing a high-throughput method, the hybridization can be analyzed simultaneously for large numbers of fragments and probes.
The resulting hybridized complex is then subjected to detection methods, such as fluorescence scanning or autoradiography, to determine the identity of the hybridized fragments. The sequence of nucleotides in the DNA or RNA molecule can be inferred by analyzing the detected signals and their corresponding complementary probes.
Sequencing by Hybridizations is a powerful technique that enables the analysis of complex mixtures of DNA or RNA fragments in a rapid and efficient manner. It has wide applications in genomics, molecular diagnostics, and research, contributing to advancements in our understanding of genetic material.
