16S rRNA sequencing for soil microbiota, replicate 3
The V3 and V4 regions were amplified using forward primers containing the sequence CCTACGGRRBGCASCAGKVRVGAAT and reverse primers containing the sequence GGACTACNVGGGTWTCTAATCC. At the same time, indexed adapters were added to the ends of the 16S rDNA amplicons to generate indexed libraries ready for downstream NGS sequencing on Illumina Miseq. PCR reactions were performed in triplicate 25 L mixture containing 2.5 L of TransStart Buffer, 2 L of dNTPs, 1 L of each primer, and 20 ng of template DNA. DNA libraries concentration were validated by Qubit 3.0 Fluorometer. Sequencing was performed using PE250 paired-end on an Illumina MiSeq instrument (Illumina, San Diego, CA, USA) by GENEWIZ.
SUB7772945
Soil_16S_S3