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Core technology Sanger dideoxy chain termination method requires a single-stranded DNA template, normal deoxynucleotidephosphates (dNTPs), DNA polymerase, and modified nucleotides (dideoxyNTPs) in vitro. These ddNTPs were fluorescently labeled for detection in automatic sequencer. Because of lacking a 3'- hydroxyl group nucleotide analogues can not format a phosphodiester bond with 5'-Phosphate group between two nucleotides, thus terminating DNA strand extension and resulting in DNA fragments of varying length. The newly synthesized and fluorescently labelled DNA fragments are separated in size by high-resolution capillary gel electrophoresis. They finally were excited by laser light and CCD detection. After reading the each fragment, the varying length of DNA sequence interpretation was combined by the computer.
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