Amplification and Sequencing of 16S rRNA

The PCR first amplification of the V3 and V4 regions of 16S rRNA was performed using 341F (5′ TCGTCGGCAGCGTC-AGATGTGTATAAGAGACAG-CCTACGGGNGGCWGCAG-3′) and 805R (5′ GTCTCGTGG GCTCGG-, AGATGTGTATAAGAGACAG-GACTACHVGGGTATCTAATCC-3′) primers, as described previously [48 (link)]. The second PCR amplification for Illumina NexTera barcodes attachment was performed. Bioanalyzer 2100 (Agilent, Palo Alto, CA, USA) was employed to evaluate quality and product size using a DNA 7500 chip. Sequencing was performed in accordance with the instructions of the Illumina MiSeq Sequencing System (Illumina Inc., SD, USA).

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Choi J., Kim J.H., Park M, & Lee H.J. (2022). Effects of Flavonoid-Rich Orange Juice Intervention on Major Depressive Disorder in Young Adults: A Randomized Controlled Trial. Nutrients, 15(1), 145.

Publication 2022

NexTera barcodes Bioanalyzer 2100 MiSeq Sequencing System

16s rrna Dna chip Primers

Corresponding Organization : Gachon University

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Variable analysis

independent variables

Primer sequences used for the PCR first amplification of the V3 and V4 regions of 16S rRNA: 341F (5′ TCGTCGGCAGCGTC-AGATGTGTATAAGAGACAG-CCTACGGGNGGCWGCAG-3′) and 805R (5′ GTCTCGTGG GCTCGG-, AGATGTGTATAAGAGACAG-GACTACHVGGGTATCTAATCC-3′)

dependent variables

Quality and product size of the PCR amplification evaluated using Bioanalyzer 2100 (Agilent, Palo Alto, CA, USA) with a DNA 7500 chip
Sequencing performed according to the instructions of the Illumina MiSeq Sequencing System (Illumina Inc., SD, USA)

control variables

Experimental protocol and primer sequences described previously in the referenced publication [48]

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