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Related Experiment Videos

Screening poly(dA/dT)- cDNAs for gene identification.

S M Wang1, S C Fears, L Zhang

  • 1Section of Hematology and Oncology, University of Chicago Medical Center, 5841 South Maryland Avenue, MC 2115, Chicago, IL 60637-1470, USA. swang1@midway.uchicago.edu

Proceedings of the National Academy of Sciences of the United States of America
|April 13, 2000
PubMed
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Long poly(dA/dT) sequences hinder gene identification by causing template loss. A new strategy, screening poly(dA/dT)(-) cDNAs, enhances gene discovery efficiency for genomic studies.

Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Many human genes remain unidentified despite extensive research efforts.
  • Long poly(dA/dT) sequences at the 3' end of cDNA templates are a significant obstacle.
  • These sequences cause random hybridization, leading to template loss during normalization/subtraction, particularly affecting low-abundance gene copies.

Purpose of the Study:

  • To address the challenge of identifying previously undiscovered genes.
  • To develop a strategy to overcome the limitations posed by poly(dA/dT) sequences in gene identification.
  • To enhance the efficiency of genome-wide gene discovery.

Main Methods:

  • Development of a strategy termed "screening poly(dA/dT)(-) cDNAs for gene identification."

Related Experiment Videos

  • Application of this strategy to identify genes that were previously lost due to poly(dA/dT) sequence interference.
  • Utilizing normalization/subtraction techniques in conjunction with the new screening method.

    • Main Results:

    • The presence of long poly(dA/dT) sequences was identified as a major cause for the loss of many cDNA templates.
    • The developed strategy effectively overcomes the obstacle of poly(dA/dT) sequences.
    • Significant enhancement in the efficiency of genome-wide gene identification was achieved.

    Conclusions:

    • The screening poly(dA/dT)(-) cDNAs strategy significantly improves gene identification efficiency.
    • This approach is crucial for advancing functional genomic studies in the post-genomic era.
    • Overcoming technical hurdles like poly(dA/dT) sequences is key to comprehensive genome exploration.