title: Bacterial community in SAMN02953206 (1014)
design: The V3-V5 fragment of the 16S rRNA gene was amplified by the polymerase chain reaction (PCR) and pyrosequenced using primers previously tested by Klindworth et al. (2013). The forward PCR primer Bakt_341F was adapted to 454 amplicon sequencing by attachment of an M13 linker (bold and underlined) to the 454 sequence adapter A (bold) and a 454 amplicon sequencing specific 4-mer amplification key (italics) followed by a 10-mer barcode sequence to aid multiplexing (NNNN): (5’- CCATCTCATCCCTGCGTGTCTCCGACTCAGNNNNCACGACGTTGTAAAACGAC -3’). The reverse template specific primer: Bakt_805R (italics) was attached to the 25-mer Lib-L specific sequence adapter B (underlined) (5’- CCTATCCCCTGTGTGCCTTGGCAGTCGACTACHVGGGTATCTAATCC-3’). molecular identifier for sample, SAMN02953206: TAGAGACGAG