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Related Experiment Video

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Methods to Discover Alternative Promoter Usage and Transcriptional Regulation of Murine Bcrp1

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Published on: May 27, 2016

Endonuclease-mediated long PCR and its application to restriction mapping.

C Her¹, R M Weinshilboum

¹Department of Pharmacology, Mayo Medical School, Mayo Clinic, Mayo Foundation, Rochester, MN 55905, USA.

Current Issues in Molecular Biology

|July 31, 2001

Summary

Long PCR (polymerase chain reaction) enables DNA amplification up to 40 kb. Novel applications simplify restriction mapping and improve amplification specificity and yield for DNA studies.

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Area of Science:

Molecular Biology
Genetics

Background:

The polymerase chain reaction (PCR) is a fundamental technique in DNA research.
Long PCR has expanded DNA amplification capabilities to fragments up to 40 kb.

Purpose of the Study:

To describe two novel applications of long PCR.
To demonstrate simplified restriction mapping using long PCR primers.
To show enhanced amplification specificity and yield through pre-amplification restriction digestion.

Main Methods:

Utilizing long PCR to amplify DNA fragments up to 40 kb.
Employing long PCR primers as probes for restriction mapping.
Applying restriction digestion before long PCR to enhance specificity.

Main Results:

Long PCR primers can serve as probes for restriction mapping of amplified DNA fragments.
Pre-amplification restriction digestion selectively suppresses amplification of homologous sequences.
Achieved enhanced specificity and yield in long PCR amplification.

Conclusions:

Two complementary long PCR techniques, combined with restriction digestion, offer significant advantages.
These methods simplify DNA analysis and improve amplification efficiency.
The described techniques have broad applicability in molecular biology laboratories performing PCR.