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The second nested primer set should only amplify the intended product from the first round of amplification and not non-specific product. This allows running more total cycles while minimizing non-specific products. This is useful for rare templates or PCR with high background.
Annealing of the 3' end of one primer to itself or the second primer may cause primer extension, resulting in the formation of so-called primer dimers, visible as low-molecular-weight bands on PCR gels. [15] Primer dimer formation often competes with formation of the DNA fragment of interest, and may be avoided using primers that are designed ...
Primer Premier is a bioinformatics software used for various PCR applications. It supports the design of degenerate primers for amplifying a related set of nucleotide sequences for the detection of common traits amongst organisms, as well as to determine heredity. [1] The software also designs tagged and nested primers for multiplex PCR ...
It involves an initial PCR with primers that have an overlap and a second PCR using the products as the template that generates the final full-length product. This technique may substitute for ligation-based assembly. [8] In colony PCR, bacterial colonies are screened directly by PCR, for example, the screen for correct DNA vector constructs ...
The primer design for all primers pairs has to be optimized so that all primer pairs can work at the same annealing temperature during PCR. Multiplex-PCR was first described in 1988 as a method to detect deletions in the dystrophin gene. [1] It has also been used with the steroid sulfatase gene. [2]
OLIGO Primer Analysis Software is a software for DNA primer design. [ 1 ] [ 2 ] The first paper describing this software was published in 1989. [ 3 ] The program is a real time PCR primer and probe search and analysis tool.
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