Metagenomic analysis of the biofilm community in the corrosive biofilm of mild steel surface
DNA Extraction : DNA extraction is done using the suitable method for the sample type from commercially available kits suchas QIAGEN, ZYMO RESEARCH, ThermoFisherDna extraction was done as per the manufacturer's recommendation.PCR Amplification of V3-V4 Region of 16s Gene:Composition of TAQ Master MIX:1) High-Fidelity DNA Polymerase2) 0.5mM dNTPs3) 3.2mM MgCl24) PCR Enzyme BufferPrimer Details:V13F:- 5 AGAGTTTGATGMTGGCTCAG3V13R:- 5 TTACCGCGGCMGCSGGCAC3Conditions Used:40ng of Extracted DNA is used for amplification along with 10pM of each primerInitial Denaturation: 95 degree C25 Cycles of the following condition:Denaturation @ 95 deg C for 15 sec, Annealing @ 60 deg C for 15 sec, Elongation @ 72 deg C for 2 minsFinal Extension at 72 deg C for 10 mins and Hold at 4 deg C.