title: 16S rRNA gene seq from Vasai soil_Untreated 2(C2)
design: PCR amplification and nanopore sequencing was done using the primers 27F 5AGAGTTTGATCMTGGCTCAG 3 and 1492R 5AAGGAGGTGATCCAGCCGCA 3. PCR was performed using the following thermal cycling conditions  initial denaturation 95C  2 min, followed by 25 cycles of denaturation 95C  30 sec, annealing 60C  30 sec, extension 72C  2 min, and a final extension 72C  10 min, then hold at 4C. Montage PCR Clean up kit (Millipore Corporation, USA) was used to remove unincorporated PCR primers and dNTPs from PCR products. The quality and quantity of the PCR product was checked using Qubit Fluorometer 3.0. Nanopore sequencing was performed with 1 g of the amplified DNA. The sequencing workflow had the following steps  end repair/dA tailing, ligation of barcode, adapter barcoding, PCR end repair/dA tailing, blunt-end adapter ligation, purification using AMPure XP bead binding, priming and loading the SpotON flow cell.