Molecular characterization of cyanobacteria isolates

For molecular characterization genomic DNA of the cyanobacteria isolate was extracted using the traditional xanthogenate technique (Tillett and Neilan, 2000). For the partial amplification of the 16S rRNA gene, a forward primer (359F, 5'-GGG GAA TYT TCC GCA ATG GG-3') and a reverse primer (781R, 5'-GAC TAC TGG GGT ATC TAA TCC CAT T-3') were used (Nübel et al., 1997). The 16S rRNA amplification was performed using 25µl aliquots containing 30–50µg DNA template, 200µM dinucleotide triphosphates, 0.4µM of forward and reverse primers, 1 U/µl Taq Polymerase and 1.5µM MgCl2, (BioRad, DNA Engine, Peltier Thermal Cycler). The reaction mixture was incubated in a Thermal cycler for DNA amplification (Singh P, et al., 2015). The amplified products were sequenced by Sanger’s method and the obtained sequence was compared with the NCBI database using BLAST tool. MEGA 11 software was used for making maximum likelihood tree for phylogenetic analysis (Guindon and Gascuel, 2003).