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Polymerase cycling assembly (or PCA, also known as Assembly PCR) is a method for the assembly of large DNA oligonucleotides from shorter fragments. The process uses the same technology as PCR, but takes advantage of DNA hybridization and annealing as well as DNA polymerase to amplify a complete sequence of DNA in a precise order based on the single stranded oligonucleotides used in the process.
A strip of eight PCR tubes, each containing a 100 μL reaction mixture Placing a strip of eight PCR tubes into a thermal cycler. The polymerase chain reaction (PCR) is a method widely used to make millions to billions of copies of a specific DNA sample rapidly, allowing scientists to amplify a very small sample of DNA (or a part of it) sufficiently to enable detailed study.
The protocols for 5' or 3' RACES differ slightly. 5' RACE-PCR begins using mRNA as a template for a first round of cDNA synthesis (or reverse transcription) reaction using an anti-sense (reverse) oligonucleotide primer that recognizes a known sequence in the middle of the gene of interest; the primer is called a gene specific primer (GSP). The ...
InterSequence-Specific PCR (or ISSR-PCR) is method for DNA fingerprinting that uses primers selected from segments repeated throughout a genome to produce a unique fingerprint of amplified product lengths. [16] The use of primers from a commonly repeated segment is called Alu-PCR, and can help amplify sequences adjacent (or between) these repeats.
Current Protocols is a series of laboratory manuals for life scientists. The first title, Current Protocols in Molecular Biology, was established in 1987 by the founding editors Frederick M. Ausubel, Roger Brent, Robert Kingston, David Moore, Jon Seidman, Kevin Struhl, and John A. Smith of the Massachusetts General Hospital Department of Molecular Biology and the Harvard Medical School ...
Scheme of touchdown PCR. The first annealing between primer and DNA occurs at the initial temperature, the second after the temperature has been lowered by Δ T {\displaystyle \Delta T} . Therefore, the product of the second annealing is disadvantaged by 2 − 1 {\displaystyle 2^{-1}} .
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