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

Screening cosmid libraries with oligonucleotides corresponding to splice-site consensus sequences

G Melmer1, M Buchwald

  • 1Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.

Genetic Analysis, Techniques and Applications
|January 1, 1994
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

Complex tools and motor-to-mechanical transformations.

Scientific reports·2022
Same author

SIAH ubiquitin ligases target the nonreceptor tyrosine kinase ACK1 for ubiquitinylation and proteasomal degradation.

Oncogene·2012
Same author

Ubiquitin conjugase UBCH8 targets active FMS-like tyrosine kinase 3 for proteasomal degradation.

Leukemia·2010
Same author

Fcgamma receptor 1 (CD64), a target beyond cancer.

Current pharmaceutical design·2009
Same author

Defective hematopoiesis and hepatic steatosis in mice with combined deficiencies of the genes encoding Fancc and Cu/Zn superoxide dismutase.

Blood·2001
Same author

Fanconi anemia group C protein prevents apoptosis in hematopoietic cells through redox regulation of GSTP1.

Nature medicine·2001

We developed a new method using splice site oligonucleotides to identify genes in genomic DNA. This technique efficiently screens cosmid libraries, confirming gene presence through hybridization.

Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • Identifying genes within large genomic DNA sequences is a significant challenge in molecular biology.
  • Traditional methods for gene identification can be time-consuming and labor-intensive.

Purpose of the Study:

  • To develop a novel method for efficient gene identification in genomic DNA.
  • To establish optimal conditions for screening cosmid libraries using specific oligonucleotide probes.

Main Methods:

  • Designing short oligonucleotides based on consensus splice site sequences.
  • Utilizing these oligonucleotides to screen cosmid libraries under specific hybridization and washing conditions.
  • Confirming gene presence by re-hybridizing isolated cosmid DNA with a different splice-site oligonucleotide.

Related Experiment Videos

Main Results:

  • The developed oligonucleotide-based method successfully identifies cosmids containing genes.
  • Specific hybridization and washing conditions were determined for effective library screening.
  • Cross-hybridization confirmed the presence of genes within the identified cosmids.

Conclusions:

  • Short oligonucleotides derived from splice sites provide a reliable method for gene identification in genomic DNA.
  • This approach offers an efficient way to screen cosmid libraries for gene-containing clones.
  • The method facilitates the discovery and characterization of genes within complex genomes.