A DNA sequencing assay wherein a single DNA polymerase enzyme is affixed at the bottom of a zero-mode waveguide (ZMW) with a single molecule of DNA as a template. The Pacific Biotechnologies version is a single-molecule real-time (SMRT) sequencing assay. The ZMW is a structure that creates an illuminated observation volume that is small enough to observe only a single nucleotide of DNA being incorporated by DNA polymerase. Each of the four DNA bases is attached to one of four different fluorescent dyes. When a nucleotide is incorporated by the DNA polymerase, the fluorescent tag is cleaved off and diffuses out of the observation area of the ZMW where its fluorescence is no longer observable. A detector detects the fluorescent signal of the nucleotide incorporation, and the base call is made according to the corresponding fluorescence of the dye.
Preferred term label
Pacific Bioscience long-read sequencing assay
Synonyms
Pac-Bio long-read sequencing assay
Textual definition
A DNA sequencing assay wherein a single DNA polymerase enzyme is affixed at the bottom of a zero-mode waveguide (ZMW) with a single molecule of DNA as a template. The Pacific Biotechnologies version is a single-molecule real-time (SMRT) sequencing assay. The ZMW is a structure that creates an illuminated observation volume that is small enough to observe only a single nucleotide of DNA being incorporated by DNA polymerase. Each of the four DNA bases is attached to one of four different fluorescent dyes. When a nucleotide is incorporated by the DNA polymerase, the fluorescent tag is cleaved off and diffuses out of the observation area of the ZMW where its fluorescence is no longer observable. A detector detects the fluorescent signal of the nucleotide incorporation, and the base call is made according to the corresponding fluorescence of the dye.
Suggested parent term
DNA sequencing assay
Attribution
0000-0002-5260-9315