Selecting Specific PCR Primers with MFEprimer

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1275)

Abstract

Selecting specific primers is crucial for polymerase chain reaction (PCR). Nonspecific primers will bind to unintended genes and result in nonspecific amplicons. MFEprimer is a program for checking the specificity of PCR primers against the background DNA. In this chapter, we introduce: (1) the factors that affect the specificity of primers; (2) the principle of MFEprimer and its settings; (3) how to use the MFEprimer to examine the specificity of primers.

Key words

PCR Primer specificity MFEprimer 
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Notes

Acknowledgement

This work was supported by the National Basic Research Project (973 program) (2012CB518200), the General Program (31371345, 30900862, 30973107, 81070741, 81172770) of the Natural Science Foundation of China, the State Key Laboratory of Proteomics of China (SKLP-O201104, SKLP-K201004, SKLP-O201002), and the Special Key Programs for Science and Technology of China (2012ZX09102301-016).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Beijing Institute of Radiation Medicine, State Key Laboratory of ProteomicsCognitive and Mental Health Research Center of PLABeijingChina

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