A primer3-based bprimer batch PCR primer design method

Abstract

The invention provides a bPrimer batch PCR primer design method based on a Primer 3.The method includes the steps that an original sequence of a target DNA sequence is obtained; a high-frequency polymorphic locus in DNA polymorphic data is extracted; the high-frequency polymorphic locus is labeled; an annotated sequence of the labeled high-frequency polymorphic locus is output, and the annotated sequence and the original sequence are identical in basic group length; the annotated sequence is read to generate candidate primers; the candidate primers are screened.The bPrimer batch PCR primer design method based on the Primer 3 can avoid the high-frequency polymorphic locus, reduces amplification failure caused by genetic diversity of target population, can detect specificity of primers in batches, reduces amplification failure caused by nonspecific amplification, primer dimer and other reasons, can be used for evaluating specificity of existing primers, and can automatic segment long target segments.

Description

technical field [0001] The present invention relates to the technical field of PCR primer design, more specifically, relates to a bPrimer batch PCR primer design method based on Primer3. Background technique [0002] Polymerase Chain Reaction (PCR), as one of the most basic molecular biology experiments, can amplify the copy number of target DNA by millions of times in vitro, so it is widely used in genetic engineering and diagnostic reagents and other fields. [0003] A key factor for the success of PCR experiments is the design of PCR primers. Therefore, in order to design better primers, a wide variety of PCR primer design software has been invented. The earliest and most basic PCR primer design software is Primer0.5, and then Primer3, an open source PCR primer design software that is now widely used, is designed on the basis of Primer0.5. The most basic functions of Primer3 include designing PCR primers and designing hybridization probes, and it has the advantages of f...

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Problems solved by technology

[0009] The purpose of the present invention is to provide a kind of bPrimer batch PCR primer design method based on Primer3, to solve the problem that the prior PCR primer design method described in the background technology causes the failure of primer design because of not preventing genetic diversity when designing PCR primers

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