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The last 10-12 bases at the 3' end of a primer are sensitive to initiation of polymerase extension and general primer stability on the template binding site. The effect of a single mismatch at these last 10 bases at the 3' end of the primer depends on its position and local structure, reducing the primer binding, selectivity, and PCR efficiency.
Sequence-context specific BLAST, more sensitive than BLAST, FASTA, and SSEARCH. Position-specific iterative version CSI-BLAST more sensitive than PSI-BLAST: Protein: Angermueller C, Biegert A, Soeding J [3] 2013 CUDASW++ GPU accelerated Smith Waterman algorithm for multiple shared-host GPUs: Protein: Liu Y, Maskell DL and Schmidt B: 2009/2010 ...
Local and NCBI Genbank BLAST search; Open reading frame finder; Restriction enzyme finder with integrated REBASE [5] restriction enzymes list; Integrated Primer3 package [6] for PCR primer design; Plasmid construction and annotation; Cloning in silico by designing of cloning vectors; Genome mapping of short reads with Bowtie, BWA, [7] and UGENE ...
In bioinformatics, BLAST (basic local alignment search tool) [3] ... To save more time, a newer version of BLAST, called BLAST2 or gapped BLAST, has been developed ...
The free NCBI tool Primer-BLAST integrates primer design and BLAST search into one application, [6] as do commercial software products such as ePrime and Beacon Designer. Computer simulations of theoretical PCR results ( Electronic PCR ) may be performed to assist in primer design by giving melting and annealing temperatures, etc. [ 7 ]
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.
A BLAST variant called MegaBLAST indexes 4 databases to speed up alignments. [9] BLAT can extend on multiple perfect and near-perfect matches (default is 2 perfect matches of length 11 for nucleotide searches and 3 perfect matches of length 4 for protein searches), while BLAST extends only when one or two matches occur close together. [1] [9]
Beacon Designer designs primers and probes for real-time PCR (polymerase chain reaction) assays.It is compatible to work on Windows as well as on Mac. The software currently supports the following real-time PCR chemistries for primer and probe design: