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Linear Amplification Mediated PCR – Localization of Genetic Elements and Characterization of Unknown Flanking DNA

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Published on: June 25, 2014

Constructing a DNA ladder Range for Lambda Phage by multiplex PCR.

R Gopalakrishnan¹, S Joseph, S Sellappa

¹Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore, Tamil Nadu, India.

Iranian Journal of Microbiology

|February 21, 2012

Summary

This summary is machine-generated.

Researchers developed a cost-effective method to create DNA ladders using multiplex PCR and lambda phage DNA. This technique amplifies specific DNA fragments, providing a valuable alternative to commercial DNA molecular weight markers.

Keywords:

DNA ladder Lambda phage DNA Multiplex PCR

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Area of Science:

Molecular Biology
Biotechnology

Background:

DNA ladders are essential tools for determining the size of unknown DNA fragments.
Various DNA ladders exist, tailored for specific DNA length ranges.
Primer design targeting conserved sequences enables the generation of DNA fragments with known sizes.

Purpose of the Study:

To describe a novel method for preparing DNA ladders.
To utilize multiplex PCR for amplifying DNA fragments of specific sizes.
To offer a cost-effective alternative to commercial DNA molecular weight markers.

Main Methods:

A DNA ladder was prepared using multiplex polymerase chain reaction (PCR).
Primers were designed based on the 1216-2136 sequence of lambda phage DNA.
Specific DNA fragments were amplified and extracted.

Main Results:

Multiplex PCR successfully amplified lambda phage DNA fragments.
One forward and six different reverse primers were used for amplification.
Successful amplification was achieved for fragments of 100, 200, 400, 600, 800, and 1000 base pairs (bp).

Conclusions:

The described method provides a cost-effective approach to DNA ladder preparation.
This technique offers a viable alternative to expensive commercial DNA molecular weight markers.