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Primer-blast tries to find target-specific primers by placing candidate primers on unique template regions that are not similar to other targets.
Go to the Primer BLAST submission form. Enter the target sequence in FASTA format or an accession number of an NCBI nucleotide sequence in the PCR Template section of the form. If the NCBI mRNA reference sequence accession number is used, the tool will automatically design primers that are specific to that splice variant. If one or both primer ...
Primer-BLAST can only find non-specific primers for your template since there are other targets that are similar or identical to your template. You may choose to re-search for specific primers by allowing some of the highly similar off-targets if they are irrelevant to your PCR experiment.
Jun 18, 2012 · Primer-BLAST allows users to design new target-specific primers in one step as well as to check the specificity of pre-existing primers. Primer-BLAST also supports placing primers based on exon/intron locations and excluding single nucleotide polymorphism (SNP) sites in primers.
Sep 22, 2023 · Primer-BLAST is a combination of Primer3, a popular primer designing tool, and NCBI BLAST. Primer-BLAST has two basic modes. Design primers to specifically amplify a template sequence or a set of related templates from a background nucleic acid population (BLAST database).
Oct 16, 2022 · The Primer-BLAST input form has four main sections: Sequence input (PCR Template and Primer Parameters) Exon / Intron selection. Primer Pair Specificity Checking Parameters (BLAST database selection and options) Advanced Parameters.
Nov 28, 2023 · The Basic Local Alignment Search Tool (BLAST) finds regions of local similarity between sequences. The program compares nucleotide or protein sequences to sequence databases and calculates the statistical significance of matches.
