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

Long PCR for VNTR analysis.

K L Richie1, M D Goldsborough, M M Darfler

  • 1Chemical Sciences and Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, MD, USA.

Journal of Forensic Sciences
|December 3, 1999
PubMed
Summary

Long PCR offers a powerful method for amplifying long DNA targets, even from degraded samples. This technique shows promise for forensic DNA analysis, though larger alleles amplify at lower levels.

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

Optimising the biocompatibility of 3D printed photopolymer constructs in vitro and in vivo.

Biomedical materials (Bristol, England)·2019
Same author

Optimization of the Promega PowerSeq™ Auto/Y system for efficient integration within a forensic DNA laboratory.

Forensic science international. Genetics·2017
Same author

Nanoscale platinum printing on insulating substrates.

Nanotechnology·2013
Same author

Printing of protein microarrays via a capillary-free fluid jetting mechanism.

Proteomics·2005
Same author

Standardization of PCR amplification for fragile X trinucleotide repeat measurements.

Clinical genetics·2002
Same author

Turkish population data with the CODIS multiplex short tandem repeat loci.

Forensic science international·2001

Area of Science:

  • Molecular Biology
  • Forensic Science
  • Genetics

Background:

  • Polymerase Chain Reaction (PCR) revolutionized DNA analysis due to its sensitivity and speed.
  • Restriction Fragment Length Polymorphism (RFLP) offers higher discrimination but requires more DNA and time.
  • Combining RFLP and PCR presents potential advantages for specific applications.

Purpose of the Study:

  • To evaluate the efficacy of Long PCR for amplifying long DNA targets (>0.5 kb to >20 kb).
  • To assess the robustness of Long PCR using degraded DNA samples.
  • To compare Long PCR performance with RFLP methods in forensic contexts.

Main Methods:

  • Utilized a Taq/Pyrococcus DNA polymerase system for Long PCR.
  • Extracted DNA from bloodstains using Chelex.

Related Experiment Videos

  • Amplified genomic DNA targets (D2S44 and D5S110 loci) and tested with partially degraded samples.

    • Main Results:

    • Successfully amplified 1-20 ng of Chelex-extracted DNA, suitable for Amp-FLP technology.
    • Demonstrated Long PCR's robustness with partially degraded blood samples compared to RFLP.
    • Detected all D2S44 and D5S110 alleles, but larger alleles amplified less efficiently than smaller ones.

    Conclusions:

    • Long PCR is a viable technique for amplifying long DNA targets from forensic samples.
    • The method shows robustness and potential for forensic DNA profiling.
    • Further optimization may be needed to improve amplification efficiency for larger alleles.