2.3 Identification of potent probionts
The biochemical characterization of the test cultures were
carried out for genus identification according to Bergey’s
Manual of Systematic Bacteriology. Further
identification of the potent probionts was achieved by 16S
rRNA sequencing and NCBI-BLAST search. The DNA was
isolated by HiPurA bacterial DNA isolation and purification
kit (Hi-media, Mumbai) and used as template. PCR
amplification of the 16S rRNA was done by using master mix
kit, MEDOX-MIX (Medox, India) as per user manual. The
primers and PCR conditions were adapted from previous
report [16]. The PCR product was subsequently sequenced and the sequence information of the isolate was compared with the
existing sequences available in the GenBank of NCBI using
BLAST search. The sequence was aligned using ClustalW
program within BioEdit version 7.0.5.3 and phylogenetic tree
was constructed by Neighbor-Joining method.