Learn DNA Primer Design for Polymerase Chain Reaction

Primer Blast, Primer Design Tools, Primer3Plus, DNA Primer Design, PCR for Beginner Course

SKU: linkshare.course.3945794 Category:

In this Bioinformatics course you will be find out how to DNA Primer Design for polymerase chain reaction. Primer BLAST performs only a specificity check when a target template and both primers are provided. Design primers for single- or multi-insert cloning or for your site-directed mutagenesis experiment (insertion, deletion, replacement) with our primer design tool. Primer3 is a computer program that suggests PCR primers for a variety of applications, for example to create STSs (sequence tagged sites) for radiation hybrid mapping, or to amplify sequences for single nucleotide polymor- phism discoveryPolymerase chain reaction (PCR) stepsDenaturingAnnealingExtensionSpecification of Primer DesignAim for the GC content to be between 40 and 60% with the 3′ of a primer ending in G or C to promote bindingA good length for PCR primers is generally around 18-30 bases. Try to make the melting temperature (Tm) of the primers between 65C and 75C, and within 5C of each otherA good length for PCR primers is generally around 18-30 bases. Specificity usually is dependent on length and annealing temperature. The shorter the primers are, the more efficiently they will bind or anneal to the target. However, a primer should not be too long 30-mer primers) or too short. Short primers produce inaccurate, nonspecific DNA amplification product, and long primers result in a slower hybridizing rate. . One also needs to avoid primer-primer annealing which creates primer dimers and disrupts the amplification process

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