16S_MiSeq_of_in_vitro_colonic_fermentation_APGH001_S098_Apple_'Monty's Surprise'_Rep3_18 h
At sampling time, 1 mL aliquots of fermenta were immediately centrifuged at 16,000 x g for 5 min at 4C, and the pellets stored at -80C. DNA was extracted from the pellet using the Qiagen DNeasy PowerLyzer PowerSoil Kit (supplied by Bio-Strategy Limited, Auckland, New Zealand), with the following modifications. For the cell lysis step, the PowerBead tubes containing the pellets (and C1 solution) were first heated at 70C for 10 min, then cooled on ice for 5 min, then shaken at 4.0 m/s for 45 s using a FastPrep-24 5G (MP Biomedicals, Ohio, USA), and cooled on ice for 5 min before further processing according to the manufacturers instructions. The DNA was dispensed into aliquots and stored at -80C until use. Preparation of amplicon libraries and sequencing of the V3-V4 regions of the 16S rRNA gene of the bacteria genomic DNA was performed at Massey Genome Service (Palmerston North, New Zealand) using a 2  250 base paired-end run on an Illumina MiSeq instrument.
SUB15490449
ExpR10773D088_APGH001_S098