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Single-molecule real-time sequencing utilizes a zero-mode waveguide (ZMW). [1] A single DNA polymerase enzyme is affixed at the bottom of a ZMW with a single molecule of DNA as a template. 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 sequencing is the process of determining the nucleotide order of a given DNA fragment. So far, most DNA sequencing has been performed using the chain termination method developed by Frederick Sanger. This technique uses sequence-specific termination of a DNA synthesis reaction using modified nucleotide substrates.
The use of DNA sequencing has also led to the development of new forensic techniques, such as DNA phenotyping, which allows investigators to predict an individual's physical characteristics based on their genetic data. In addition to its applications in forensic science, DNA sequencing has also been used in medical research and diagnosis.
In molecular biology, and more importantly high-throughput DNA sequencing, a chimera is a single DNA sequence originating when multiple transcripts or DNA sequences get joined. Chimeras can be considered artifacts and be filtered out from the data during processing [ 1 ] to prevent spurious inferences of biological variation. [ 2 ]
Single-cell DNA genome sequencing involves isolating a single cell, amplifying the whole genome or region of interest, constructing sequencing libraries, and then applying next-generation DNA sequencing (for example Illumina, Ion Torrent). Single-cell DNA sequencing has been widely applied in mammalian systems to study normal physiology and ...
Nanopore sequencing is a third generation [1] approach used in the sequencing of biopolymers — specifically, polynucleotides in the form of DNA or RNA. Nanopore sequencing allows a single molecule of DNA or RNA be sequenced without PCR amplification or chemical labeling.
Single molecule fluorescent sequencing is one method of DNA sequencing. The core principle is the imaging of individual fluorophore molecules, each corresponding to one base. [1] By working on single molecule level, amplification of DNA is not required, avoiding amplification bias. The method lends itself to parallelization by probing many ...
A distinct group of DNA-binding proteins is the DNA-binding proteins that specifically bind single-stranded DNA. In humans, replication protein A is the best-understood member of this family and is used in processes where the double helix is separated, including DNA replication, recombination, and DNA repair. [ 123 ]