infrastruktura: Polymerase chain reaction

Evidence: 100 • authority: 72

The polymerase chain reaction (PCR) is a laboratory method widely used to amplify copies of specific DNA sequences rapidly, to enable detailed study. PCR was invented in 1983 by American biochemist Kary Mullis at Cetus Corporation. Mullis and biochemist Michael Smith, who had developed other essential ways of manipulating DNA, were jointly awarded the Nobel Prize in Chemistry in 1993. PCR is fundamental to many of the procedures used in genetic testing, research, including analysis of ancient samples of DNA, and identification of infectious agents. Using PCR, copies of very small amounts of DNA sequences are exponentially amplified in a series of cycles of temperature changes. PCR is now a…

Evidence: 56 • authority: 72

Gel electrophoresis is a method for separation and analysis of biomacromolecules (DNA, RNA, proteins, etc.) and their fragments, based on their size and charge as they move through a gel under an electric field. It is used in clinical chemistry to separate proteins by charge or size and in biochemistry and molecular biology to separate a mixed population of DNA and RNA fragments by length, to estimate the size of DNA and RNA fragments, or to separate proteins by charge. Nucleic acid molecules are separated by applying an electric field to move the negatively charged molecules through a gel matrix of agarose, polyacrylamide, or other substances. Shorter molecules move faster and migrate fart…

Evidence: 37 • authority: 80

POLYMERASE CHAIN REACTION (PCR) AND REAL TIME PCR (For Detection and Quantification of Nucleic Acid)

Evidence: 37 • authority: 80

POLYMERASE CHAIN REACTION (PCR) AND REAL-TIME PCR (For Detection and Quantification of Nucleic Acid)

Evidence: 37 • authority: 88

2-step Polymerase Chain Reaction (PCR) v1. Polymerase Chain Reaction (PCR) is a method of making multiple copies of a DNA sequence, involving repeated reactions with a polymerase enzyme, which comes in different types, such as Taq-polymerase, Phusion or Q5. To run a PCR reaction, one needs a DNA template to be amplified, one set of forward and reverse primers, a polymerase enzyme as well as various buffers. This protocol describes the procedure to perform a two-step PCR reaction, in which the primers contain an extra non-binding overlap sequence which allows the assembly of a new sequence from a specific region of the template DNA. This is done by using two sets of primers, one which adds a…