oral metagenome of mus musculus
Bacteria were obtained from the teeth and gingival surface, or from the ligature surface, by swab collection and bacteria were eluted in PBS in cell lysis buffer from a DNeasy kit (Qiagen, Valencia, CA, USA) as described (Grice et al., 2010a). After a 60-s vortex, DNA present in the buffer was isolated with the DNeasy kit and quantitated with a spectrophotometer (Tecan, Mannedorf, Switzerland). 515F/806R primer with golay barcode in the reverse primer was used to amplify the V4 region of 16S ribosomal DNA (16S rDNA; IDT, Coralville, IA, USA) following the procedure described by Grice et al to obtain bacteria from db/db diabetic mice (Grice et al., 2010a). PCR was performed in 50-uL reactions and then purified with both Agencourt XP DNA purification beads (Beckman Coulter, Beverly, MA, USA) and QiaAmp DNA Mini Spin Columns (Qiagen).
SUB2771100
NG3.sb.cbc124