Metagenome: oil contaminated barley planted soil, replicate 1
The 16S rRNA gene variable region V4 PCR primers 515/806 with barcode on the forward primer were used in a 30 cycle PCR  (5 cycle used on PCR products) using the HotStarTaq Plus Master Mix Kit (Qiagen, USA) under the following conditions: 94 C for 3 minutes, followed by 28 cycles of 94 C for 30 seconds, 53 C for 40 seconds and 72 C for 1 minute, after which a final elongation step at 72 C for 5 minutes was performed. After amplification, the PCR products were checked in 2% agarose gel to determine the success of amplification and the relative intensity of bands. Multiple samples were pooled together in equal proportions based on their molecular weight and DNA concentrations. Pooled samples were purified using calibrated Ampure XP beads. Then the pooled and purified PCR product was used to prepare DNA library by following Illumina TruSeq DNA library preparation protocol.
Kuwait Institute for Scientific Research (KISR)
BO_R1