A Comparative analysis of Long PCR and Standard PCR technique in detecting the Wolbachia Endosymbiont

Order of Publishing in Issue: 
11
Volume :6
Issue :4
October, 2012
Page No: 
472-478
Authors: 
Sumithra [1], N. M. Guruprasad [2] and H. P. Puttaraju [1]*
Address: 
[1] Department of Biological Sciences, Bangalore University, Bangalore - 560056, India
Address: 
[2] National Bureau of Agriculturally Important Insects (NBAII) Bangalore-560024, India

Abstract

Allele specific polymerase chain reaction (Standard PCR) is being widely used to amplify Wolbachia DNA from arthropods. Preliminary tests with Wolbachia allele-specific polymerase chain reaction suggested that the assays were prone to false negative results in insects. Standard PCR frequently produced false negative results, perhaps due to low titer Wolbachia DNA mixed with host genomic DNA. On the other hand, another PCR protocol, Long PCR which uses proof reading enzyme consistently amplified Wolbachia DNA and revealed that 49% of 35 arthropod species tested positive for Wolbachia which is considerably higher than the rate of 31% obtained in Standard PCR. An attempt has been made in this study to compare both Standard and Long PCR and to evaluate Long PCR as a technique for amplifying Wolbachia DNA from a diverse array of arthropods.

Keywords: 
Polymerase Chain Reaction (PCR), Long PCR, Standard PCR, Wolbachia, Proof reading, Insects.
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