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Secondary structures in the DNA can result in folding or knotting of DNA template or primers, leading to decreased product yield or failure of the reaction. Hairpins, which consist of internal folds caused by base-pairing between nucleotides in inverted repeats within single-stranded DNA, are common secondary structures and may result in failed PCRs.
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.
The design of appropriate short or long primer pairs is only one goal of PCR product prediction. Other information provided by in silico PCR tools may include determining primer location, orientation, length of each amplicon, simulation of electrophoretic mobility, identification of open reading frames, and links to other web resources. [7] [8] [9]
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]
And a primer sequence at the end, it is a sequence whose design varies and is what will allow the design of primers and subsequent fragment amplification. In addition, one of the parts of the probe usually contains a stuffer between the target sequence and the primer sequence. The use of different stuffers allows the identification of probes ...
The 5 prime portion of the RC probe contains the reverse complement sequence of the desired target specific primer sequence. In RC-PCR, no target specific primers are present in the reaction mixture. Instead target specific primers are formed as the reaction proceeds. A typical reaction employing the approach requires four oligonucleotides. The ...
All of the basic information regarding the target is necessary here, including the gene symbol, the accession database number for the sequence in question, the length of the sequence being amplified, information about the specificity screen used such as BLAST, what splicing variants exist for the sequence, and where the exon or intron for each ...
MLST databases contain the reference allele sequences and sequence types for each organism, and also isolate epidemiological data. The websites contain interrogation and analysis software which allow users to query their allele sequences and sequence types. MLST is widely used as a tool for researchers and public healthcare workers.