Using Web Primer: Sequencing

Primer composition

Primers that contain a skewed AT/GC ratio can fail to give high specificity, or yield primers that are in other ways not well behaved. The user is allowed to enter minimum, optimal, and maximum values for the percentage of basepairs which are either G or C.

Primer Annealing

Primers also tend to dimerize and anneal to themselves. One method for accounting for this problem was developed by Hillier and Green, PCR Method. Applic., 1; 124-8, 1991. Maximum values annealing between primers are able to be set by the user.

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