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RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
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Genome-wide Surveillance of Transcription Errors in Eukaryotic Organisms
09:30

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Published on: September 13, 2018

Normalizing cDNA libraries.

Ekaterina A Bogdanov1, Irina Shagina, Ekaterina V Barsova

  • 1Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.

Current Protocols in Molecular Biology
|April 8, 2010
PubMed
Summary
This summary is machine-generated.

Characterizing rare messages in cDNA libraries is challenging due to transcript abundance variations. Duplex-specific nuclease (DSN) normalization rapidly equalizes cDNA, enabling efficient analysis of rare transcripts and full-length cDNA libraries.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Transcript abundance varies significantly in biological samples, complicating the study of rare messages.
  • Analyzing rare transcripts requires methods to overcome the dominance of highly abundant ones.

Purpose of the Study:

  • To present a method for cDNA normalization using duplex-specific nuclease (DSN).
  • To enable faster and more reliable equalization of cDNA for downstream applications.

Main Methods:

  • Utilizing duplex-specific nuclease (DSN) for cDNA normalization.
  • Generating normalized, full-length-enriched cDNA libraries.

Main Results:

  • DSN-based normalization effectively reduces the prevalence of abundant transcripts.
  • Facilitates the assessment and characterization of rare transcripts.

Conclusions:

  • DSN normalization is a fast and reliable method for cDNA equalization.
  • Enables efficient RNA analyses and the creation of normalized cDNA libraries.