title: Full 16S rRNA for anerobic microbiome
design: the whole 16S gene was amplified using the 27f (AGRGTTYGATYMTGGCTCAG) and 1492r(RGYTACCTTGTTACGACTT) primers with barcodes and adapters. 2ng DNA was used as the PCR tempalte for each sample. The PCR was run with 25 cycles following the conditions mentioned in the protocol. After that, the PCR product of each reaction underwent clean up with Select-a-Size DNA Clean & Concentrator MagBead Kit (Zymo Research, Irvine, CA) keeping fragments >300bp. The library of each reaction is then quantified using NanoDrop and pooled together with equal DNA mass. The pooled library was prepared for PacBio sequencing using the SMRTbell prep kit 3.0