Sanger Sequencing Reagents

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Generation sequencing, also known as Sanger sequencing, is a technology invented by Professor Sanger in 1975 used to determine the sequence of DNA. It uses the chain termination method, “dideoxy termination method,” or the “Sanger method.” Sanger invented this method and first determined the genome sequence of the Φ-X174 phage.

By partially labeling the four bases of ATCG with isotopes, the positions of these four bases can be determined by radiographic imaging after electrophoresis. For example, for the sequence ATCGATCGATCG, A is used as a termination marker. Three types of fragments can be obtained, namely A, ATCGA, ATCGATCGA. So the analysis knows that the base A occurs in the 1st, 5th and 9th positions. Similarly, if other bases are marked with the same method, the positions of the remaining bases can also be determined, and finally the base sequence of the sequence to be tested can be obtained.

The leading instrument for next-generation sequencing is the 3730XL from ABI. The general sequencing length can reach 1000bp. Its main principle is to use fluorescently labeled nucleotides for sequencing while synthesizing in PCR. Since the fluorescent labels of the four nucleotides are different, the sequence information is known.

As the sequencing method with the highest sequencing accuracy at present, Sanger sequencing is mainly used in a small number of individual-level studies such as routine clone detection, single-gene detection, and small-throughput screening, and is known as the "gold standard for sequencing."