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In silico PCR (CAT#: STEM-MB-0227-WXH)

Introduction

in silico PCR refers to computational tools used to calculate theoretical polymerase chain reaction (PCR) results using a given set of primers (probes) to amplify DNA sequences from a sequenced genome or transcriptome. In silico PCR analysis is a useful and efficient complementary method to ensure primer specificity for an extensive range of PCR applications from gene discovery, molecular diagnosis, and pathogen detection to forensic DNA typing.




Principle

In silico PCR refers to a virtual PCR executed by a computer program with an input of a pair or a batch of primers against an intended genome that is stored in a silicon media (e.g., a database server). In silico PCR aims to test PCR specificity including the target location and amplicon size in one or multiple target genome(s). It can identify the mismatches in primer binding sites due to known single nucleotide polymorphisms (SNPs) and/or unwanted amplicons from a homologous gene or a pseudogene. With the development of sequencing technology and rapid cost reduction, many genomes have been sequenced and annotated in databases. Such a wealth of genomic information makes in silico PCR possible.

Applications

• Optimize the design of primers for target DNA or cDNA sequences.
• Information provided by in silico PCR tools may include determining primer location, orientation, length of each amplicon, simulation of electrophoretic mobility, identification of open reading frames, and links to other web resources.

Procedure

General procedure:
1. Denaturation
2. Annealing
3. Extension