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Amplified fragment length polymorphism (AFLP-PCR or AFLP) is a PCR-based tool used in genetics research, DNA fingerprinting, and in the practice of genetic engineering. Developed in the early 1990s by Pieter Vos, [1] AFLP uses restriction enzymes to digest genomic DNA, followed by ligation of adaptors to the sticky ends of the restriction ...
The DNA sequencing is done on a chip that contains many ZMWs. Inside each ZMW, a single active DNA polymerase with a single molecule of single stranded DNA template is immobilized to the bottom through which light can penetrate and create a visualization chamber that allows monitoring of the activity of the DNA polymerase at a single molecule level.
C 0 t analysis, a technique based on the principles of DNA reassociation kinetics, is a biochemical technique that measures how much repetitive DNA is in a DNA sample such as a genome. [1] It is used to study genome structure and organization and has also been used to simplify the sequencing of genomes that contain large amounts of repetitive ...
Then, like other polymerase chain reaction processes, the DNA is amplified by the thermostable DNA polymerase: A target region with an internal section of known sequence and unknown flanking regions is identified; Genomic DNA is digested into fragments of a few kilobases by a usually low-moderate frequency (6-8 base) cutting restriction enzyme.
The process of extensive BAC library creation and tiling path selection, however, make hierarchical shotgun sequencing slow and labor-intensive. Now that the technology is available and the reliability of the data demonstrated, [14] the speed and cost efficiency of whole-genome shotgun sequencing has made it the primary method for genome ...
The DNA fragments are transferred out of the gel or matrix onto a solid membrane, which is then exposed to a DNA probe labeled with a radioactive, fluorescent, or chemical tag. The tag allows any DNA fragments containing complementary sequences with the DNA probe sequence to be visualized within the Southern blot. [1]
This method is deployed for DNA sequencing, genome walking, and DNA footprinting. [12] A related technique is amplified fragment length polymorphism, which generates diagnostic fragments of a genome. Methylation-specific PCR (MSP) is used to identify patterns of DNA methylation at cytosine-guanine (CpG) islands in genomic DNA. [13]
During sequencing, each base in the template is sequenced twice, and the resulting data are decoded according to this scheme. SOLiD (Sequencing by Oligonucleotide Ligation and Detection) is a next-generation DNA sequencing technology developed by Life Technologies and has been commercially available since 2006.