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

Simple strategy for sequencing cDNA clones

L W Zeng1, M Kreitman

  • 1Department of Ecology and Evolution, University of Chicago, IL 60636, USA. lzeng@midway.uchicago.edu

Biotechniques
|September 1, 1996
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

[A case of peripheral T-cell lymphoma with lung involvement].

Zhonghua jie he he hu xi za zhi = Zhonghua jiehe he huxi zazhi = Chinese journal of tuberculosis and respiratory diseases·2025
Same author

Evaluation of the effectiveness of air pollution control measures in Hong Kong.

Environmental pollution (Barking, Essex : 1987)·2016
Same author

Is mitochondrial DNA a strictly neutral marker?

Trends in ecology & evolution·2011
Same author

Distribution of genetic variation within and among local populations of Arabidopsis thaliana over its species range.

Molecular ecology·2006
Same author

Embryonic enhancers in the dpp disk region regulate a second round of Dpp signaling from the dorsal ectoderm to the mesoderm that represses Zfh-1 expression in a subset of pericardial cells.

Developmental biology·2003
Same author

Fitness costs of R-gene-mediated resistance in Arabidopsis thaliana.

Nature·2003
Same journal

Investigating the interactomic landscape of survival motor neuron (SMN) and the SMNΔ7 truncated protein.

BioTechniques·2026
Same journal

Antigen retrieval-immunofluorescence on free floating sections to visualize the liver lobule and its cellular makeup.

BioTechniques·2026
Same journal

Special approach of droplet digital polymerase chain reaction (ddPCR) for transgene stability of a Chinese hamster ovary (CHO) cell line.

BioTechniques·2026
Same journal

Strand-specific quantification of L1 ORF0 and related transcripts by multiplex reverse transcription with tagged primers.

BioTechniques·2026
Same journal

Why and when should we choose digital PCR?

BioTechniques·2026
Same journal

Quantitative and unbiased lung alveolar septum assessment in an LPS experimental mouse model using 2D-spatial correlation image analysis from hematoxylin and eosin slides.

BioTechniques·2026
See all related articles

Researchers developed a straightforward method for creating nested deletion subclones from cDNA. This technique simplifies DNA sequencing, reduces redundancy, and is suitable for automated sequencing platforms.

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Sequencing large cDNA clones efficiently remains a challenge.
  • Traditional methods like shotgun cloning can be redundant and labor-intensive.

Purpose of the Study:

  • To present a novel, simplified method for constructing overlapping nested deletions from cDNA clones.
  • To enable efficient and accurate sequencing of large cDNA inserts.

Main Methods:

  • Partial digestion of PCR-amplified cDNA inserts with restriction enzymes.
  • Generation of overlapping nested subfragments.
  • Recloning into plasmid vectors and screening using colony PCR.

Main Results:

  • Successfully generated nested deletion sets from cDNA clones.

Related Experiment Videos

  • Eliminated sequencing redundancy issues associated with shotgun cloning.
  • Sequenced five cDNA clones from Drosophila subobscura genes using universal primers.

    • Conclusions:

    • The described method offers a simple, single-step cloning approach for generating nested deletions.
    • This technique is efficient for sequencing large DNA fragments and is compatible with automated sequencing.
    • The method significantly improves the process of cDNA clone sequencing.