[5] Reverse Cloning Procedure for Generation of Subclones for DNA Sequencing

  • Liu Z
  • Hackett P
  • 2

    Readers

    Mendeley users who have this article in their library.
  • 0

    Citations

    Citations of this article.

Abstract

This chapter describes an easy, efficient, and reliable method for preparing the subclones of long DNA fragments for sequencing. Reverse cloning is simple; it avoids gel purification or fractionation of DNA fragments, use of linkers, and requires only a standard primer conventionally used for DNA sequencing. Repeated sequencing is minimized, thereby shortening the time of sequencing projects. The reverse cloning procedure depends on the rapid speed of the enzyme. However, a consequence of the high speed of polymerization is the lack of proofreading by the DNA polymerase. The sequencing of the both strands of a specific region of DNA, from independently generated clones, provides the necessary check on the fidelity of the overall DNA sequence. The separate generation of complementary subclones should identify any low-probability errors in subclone sequences. © 1993, Elsevier Inc. All rights reserved.

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

  • Zhanjiang Liu

  • Perry B. Hackett

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free