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cDNA amplification using one-sided (anchored) PCR.

R L Dorit¹, O Ohara

¹Yale University, New Haven, Connecticut, USA.

Current Protocols in Immunology

|April 25, 2008

Summary

This summary is machine-generated.

Anchored PCR amplifies full-length mRNA using limited sequence data. This technique employs oligo(dT) primers for targeted amplification, enabling direct sequencing or cloning of mRNA molecules.

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Amplifying full-length mRNA is crucial for gene expression studies.
Limited sequence information often hinders traditional mRNA amplification methods.

Purpose of the Study:

To present anchored PCR, a modified PCR technique for full-length mRNA amplification.
To enable amplification using minimal known mRNA sequence data.

Main Methods:

Anchored PCR utilizes oligo(dT) primers for amplification.
Primers bind to either the poly(A) tail or an enzymatically added homopolymer tail.
Two rounds of PCR amplification are performed.

Main Results:

Achieved amplification of full-length mRNA from limited sequence information.
Generated a single, amplicon product suitable for downstream applications.

Conclusions:

Anchored PCR is an effective method for full-length mRNA amplification.
The technique facilitates direct sequencing or cloning of amplified mRNA.