siqoriginal.e12_GAGTCTCA.GTGCCAGCMGCCGCGGTAA.fastq.e12_GAGTCTCA.GTGCCAGCMGCCGCGGTAA
siqoriginal.e12_GAGTCTCA.GTGCCAGCMGCCGCGGTAA.fastq.e12_GAGTCTCA.GTGCCAGCMGCCGCGGTAA
"The 16S rRNA gene V4 variable region PCR primers 515/806 (CAPORASO et al., 2011) 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: 94oC for 3 minutes, followed by 28 cycles of 94oC for 30 seconds, 53oC for 40 seconds and 72oC for 1 minute, after which a final elongation step at 72oC for 5 minutes was performed.  After amplification, PCR products are checked in 2% agarose gel to determine the success of amplification and the relative intensity of bands. Multiple samples are pooled together in equal proportions based on their molecular weight and DNA concentrations. Pooled samples are purified using calibrated Ampure XP beads. Then the pooled and purified PCR product is used to prepare DNA library by following Illumina TruSeq DNA library preparation protocol. Paired-end sequencing was performed at MR DNA (www.mrdnalab.com, Shallowater, TX, USA) on a MiSeq following the manufacturer's guidelines. The sequences from each end were joined after quality q25 trimming of the ends with MR DNA pipeline. "
SUB1547922
siqoriginal.e12_GAGTCTCA.GTGCCAGCMGCCGCGGTAA.fastq.e12_GAGTCTCA.GTGCCAGCMGCCGCGGTAA