MiSeq of Apis mellifera gut: Queen Mouthparts
The V6 V8 variable region of the 16S rRNA gene was amplified using PCR primers 799F (acCMGGATTAGATACCCKG + barcode) and bac1193R (CRTCCMCACCTTCCTC). Amplification was performed using the HotStarTaq Plus Master Mix Kit (Qiagen, USA) under the following conditions: 94  C for 3 min, followed by 28 cycles of 94  C for 30 s, 53  C for 40 s and 72  C for 1 min, with a final elongation step at 72  C for 5 min. PCR products were confirmed using a 2% agarose gel. PCR products were then used to prepare DNA libraries following Illumina MiSeq DNA library preparation protocol. Sequencing was performed at the University of Arizona Genetics Core on a MiSeq following the manufacturer s guidelines.