Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Differential display probes for cDNA arrays.

T Trenkle1, J Welsh, M McClelland

  • 1Sidney Kimmel Cancer Center, San Diego, CA, USA.

Biotechniques
|September 18, 1999
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Ecotin protects <i>Salmonella</i> Typhimurium against the microbicidal activity of host proteases.

bioRxiv : the preprint server for biology·2024
Same author

Discretion in decision to receive COVID-19 vaccines and associated socio-economic inequalities in rates of uptake: a whole-of-population data linkage study from Australia.

Public health·2023
Same author

Inequalities in life expectancy in Australia according to education level: a whole-of-population record linkage study.

International journal for equity in health·2021
Same author

YeiE Regulates Motility and Gut Colonization in Salmonella enterica Serotype Typhimurium.

mBio·2021
Same author

Space Medicine: Why Do Recently Published Papers about Telomere Length Alterations Increase our Uncertainty Rather than Reduce it?

Journal of biomedical physics & engineering·2021
Same author

Identifying long-term psychological distress from single measures: evidence from a nationally representative longitudinal survey of the Australian population.

BMC medical research methodology·2020

This study introduces a novel method using PCR-generated cDNA array probes, significantly reducing RNA requirements. This technique efficiently identifies differentially regulated genes, offering a powerful tool for molecular biology research.

Area of Science:

  • Molecular Biology
  • Gene Expression Analysis
  • Genomics

Background:

  • Traditional array hybridization methods require substantial RNA amounts.
  • Differential Display (DD) PCR is a technique for analyzing gene expression.

Purpose of the Study:

  • To develop an efficient probe generation method for cDNA arrays using DD PCR.
  • To reduce RNA input and increase the identification rate of differentially regulated genes.

Main Methods:

  • Utilized PCR with arbitrary and oligo(dT) anchor primers to generate probes.
  • Adapted the DD protocol to produce sufficient PCR product mass for array probing.
  • Employed E. coli colony arrays of expressed sequence tag (EST) clones.

Main Results:

Related Experiment Videos

  • The method requires less than 1/200 of the RNA compared to other array methods.
  • Each probe detected approximately 5% of transcribed mRNAs, sampled independently of abundance.
  • Successfully identified four novel and three known EGF-regulated genes.

Conclusions:

  • PCR-generated DD fingerprints serve as effective probes for cDNA arrays.
  • This approach enhances the identification of differentially regulated genes and minimizes the need for cloning and sequencing.
  • Archived DD fingerprints can be repurposed as probes for further data extraction.