MCK3_2
16S-Seq of rhizospheric soil: MCK3_2
High purity genomic DNA was isolated from maize rhizosphere soil samples using the E.Z.N.A.  Soil DNA Kit (Omega Bio-TEK, U.S.A.) according to the manufacturers instructions. The final DNA concentration and purification were determined by NanoDrop 2000 UV-vis spectrophotometer (Thermo Scientific, Wilmington, USA), and DNA quality was checked by 1% agarose gel electrophoresis. The V3-V4 hypervariable regions of the bacterial 16S rRNA gene were amplified with primers 338F (5-ACTCCTACGGGAGGCAGCAG-3) and 806R (5-GGACTACHVGGGTWTCTAAT-3) by thermocycler PCR system (GeneAmp 9700, ABI, USA). The PCR reactions were conducted using the following program: 3 min of denaturation at 95C, 27 cycles of 30 s at 95C, 30s for annealing at 55C, and 45s for elongation at 72C, and a final extension at 72C for 10 min. PCR reactions were performed in triplicate 20 L mixture containing 4 L of 5FastPfu buffer, 2 L of 2.5 mM dNTPs, 0.8 L of each primer (5 M), 0.4 L of FastPfu polymerase and 10 ng of template DNA. The resulted PCR products were subjected to an electrophoresis on a 2% agarose gel, and then extracted and further purified using the AxyPrep DNA Gel Extraction Kit (Axygen Biosciences, Union City, CA, USA) and quantified using QuantiFluor-ST (Promega, USA) according to the product manual. Purified amplicons were pooled in equimolar and paired-end sequenced (2  300) on an Illumina MiSeq platform (Illumina, San Diego, USA) according to the standard protocols by Majorbio Bio-Pharm Technology Co. Ltd. (Shanghai, China).
MCK3_2
SUB6027886
MCK3_2