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Related Experiment Videos

Double-stranded DNA sequence analysis.

Sherie L Morrison1

  • 1University of California Los Angeles, Los Angeles, California.

Current Protocols in Immunology
|April 25, 2008
PubMed
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This study details DNA sequencing methods for analyzing immunoglobulin variable (V) regions. The described protocols use the Sequenase Version 2.0 kit for accurate DNA sequence determination.

Area of Science:

  • Molecular Biology
  • Immunogenetics
  • Biochemistry

Background:

  • Analyzing cloned DNA sequences is crucial in molecular biology.
  • Immunoglobulin variable (V) regions require precise sequencing for genetic studies.

Purpose of the Study:

  • To provide detailed protocols for sequencing cloned immunoglobulin V regions.
  • To outline the use of the Sequenase Version 2.0 kit for DNA analysis.

Main Methods:

  • Double-stranded DNA sequencing using the dideoxy method.
  • Utilizing a modified T7 DNA polymerase (Sequenase Version 2.0 kit).
  • Performing sequencing reactions, gel electrophoresis, and analysis.

Main Results:

  • The double-stranded DNA sequencing method provides sufficient information for accurate sequence determination of immunoglobulin V regions.

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  • Protocols are established for both performing the sequencing reaction and preparing the sequencing gel.

    • Conclusions:

    • Double-stranded DNA sequencing is a reliable and accessible method for analyzing immunoglobulin V regions.
    • The Sequenase Version 2.0 kit facilitates efficient and accurate DNA sequencing.