Current Trends in Biotechnology and Pharmacy
Open Access
SCOPUS
  • Year: 2012
  • Volume: 6
  • Issue: 4

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

  • Author:
  • Sumithra 1, N. M. Guruprasad2, H. P. Puttaraju1,
  • Total Page Count: 7
  • Page Number: 472 to 478

1Department of Biological Sciences, Bangalore University, Bangalore - 560056, India

2National Bureau of Agriculturally Important Insects (NBAII), Bangalore-560024, India

*For Correspondence - puttarajuhp@hotmail.com

Online published on 29 April, 2013.

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