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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.
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 ...
Droplet Digital PCR (ddPCR) is a method of dPCR in which a 20 microliter sample reaction including assay primers and either Taqman probes or an intercalating dye, is divided into ~20,000 nanoliter-sized oil droplets through a water-oil emulsion technique, thermocycled to endpoint in a 96-well PCR plate, and fluorescence amplitude read for all ...
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.
PCR works readily with a DNA template of up to two to three thousand base pairs in length. However, above this size, product yields often decrease, as with increasing length stochastic effects such as premature termination by the polymerase begin to affect the efficiency of the PCR. It is possible to amplify larger pieces of up to 50,000 base ...
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.
Hot start PCR is a method which prevents DNA polymerase extension at lower temperature to prevent non-specific binding to minimise yield loss. Hot start PCR reduces the amount of non-specific binding through limiting reagents until the heating steps of PCR – limit the reaction early by limiting Taq DNA polymerase in a reaction.
Topoisomerase-based cloning (TOPO cloning) is a molecular biology technique in which DNA fragments are cloned into specific vectors without the requirement for DNA ligases.Taq polymerase has a nontemplate-dependent terminal transferase activity that adds a single deoxyadenosine (A) to the 3'-end of the PCR products.