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Multiplex PCR (CAT#: STEM-MB-0193-WXH)

Introduction

Multiplex PCR is the simultaneous detection of multiple targets in a single reaction well, with a different pair of primers for each target. This technique requires two or more probes that can be distinguished from each other and detected simultaneously.<br />Advantages of using multiplex PCR include:<br />More information with less sample<br />Higher throughput<br />Cost effective — fewer dNTPs, enzymes, and other consumables<br />Time saving<br />Less input material required<br />More data from limited starting materials<br />Increased accuracy of data normalization<br />Fewer pipetting errors




Principle

The basic principle of multiplex PCR is the same as that of the conventional PCR, except that more than one pair of primers are required in the same reaction. The primers can specifically combine with their corresponding DNA template, and more than one DNA fragment will be amplified in one reaction simultaneously.

Applications

• Life science research
• Clinical diagnostics
• Forensic laboratories
• Pathogen detection
• GMO (genetically modified organism) detection
• Forensic studies
• Food analysis
• Mutation and polymorphism analysis
• Gene deletion analysis
• Template quantitation
• Linkage analysis
• RNA detection

Procedure

1.Choose primer sequences
2.Test/align primer sequences
3.Single locus PCR: Initial program
4.Multiplex PCR: equimolar primer mix

Materials

Primers: Three rounds of PCR with three specific (SP) primers that have a high Tm from the known sequences and four arbitrary degenerate (AD) primers that have a low Tm for binding to the unknown sequences