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16S rDNA Identification and molecular characterization of xanthomonadaceae nml 03-0222 bacterial isolates from agricultural soil of jharkhand

Author: 
Mukesh Nitin, Rajnish Kumar, Agatha S. Khalko, Ruma Sinha, M. S. Anantha and Yogesh Kumar
Subject Area: 
Life Sciences
Abstract: 

The traditional method for identification of bacterial species depends upon different biochemical testing and colonial morphology study. Biochemical tests are simple to execute, require minimal equipment, and generally accurately differentiate between the more common species. However, they are time-consuming and results delay in final identification due to long incubation time period. Several genes common to all bacteria have been studied in sequence-based identification. The objective o present study is to isolate, identify and characterize the bacteria from agricultural farm field of Jharkhand. The use of 16S rDNA gene sequence analysis method, is considered as recent standard approach for identification for all bacteria and is widely recognized as a rapid and accurate method for identification of novel bacterial culture. Fragment of 16S rDNA gene of the DNA was isolated from the bacterial species and was amplified by PCR. DNA sequencing reaction of purified PCR amplicon was carried out with (16sF) and (16sR) forward and reverse primers. Basic Local Alignment Search Tool (BLAST) was performed and the evolutionary distances were computed using the Kimura 2-parameter method and expressed in the units of the number of base substitutions per site. Phylogenetic analyses were conducted in (MEGA 4.0) software for 1447 nt contig region of sample G. It was observed to be homologous with sequence of Genbank having accession no. EU313791.1. and was identified as Xanthomonadaceae NML 03-0222.

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