A simple and practical method that prepares high molecular weight DNA ladders

Mol Med Rep. 2012 Nov;6(5):1211-3. doi: 10.3892/mmr.2012.1061. Epub 2012 Sep 4.

Abstract

The purpose of the current study was to report a simple and practical method to prepare high molecular weight (mw) DNA ladders. The method involves 1,000-4,000-base pairs (bp) DNA fragments being amplified by polymerase chain reaction (PCR), using λ DNA as a template. The constructed plasmids are digested by restriction endonucleases to produce 5-, 6-, 8- and 10-kb DNA fragments, followed by purification and precipitation with ethanol, and mixed proportionally. The 1,000-4,000-bp DNA fragments were successfully generated by PCR and 5-, 6-, 8- and 10-kb DNA fragments were obtained through the digestion of the plasmids. The bands of the prepared high mw DNA ladder were clear and may aid future molecular biology studies.

Keywords: DNA ladder; polymerase chain reaction; restriction endonuclease digestion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry*
  • DNA / metabolism
  • DNA Fragmentation
  • DNA Restriction Enzymes / metabolism
  • Molecular Weight
  • Plasmids / metabolism
  • Polymerase Chain Reaction

Substances

  • DNA
  • DNA Restriction Enzymes