Guana site 1 (maternal nest surface A sand)
mixed culture metagenome
isolation_source
sand
collection_date
2020-08-09
geo_loc_name
USA: Northeast Florida
lat_lon
30.1207 N 81.3458 W
samp_collect_device
50 ml falcon tubes
samp_mat_process
10 ml sand plus 20 ml 1X TE or 10 ml sand plus 15 ml 1X TE-IDTE pH 8.0 1x TE Solution was added to each individual sand sample in 50 ml falcon tubes. Samples were shaken gently by hand then set on a rocking platform for 1 hour at room temperature, shaking gently by hand every 15 minutes. Samples rested until sand sunk to bottom of tube then the surface buffer was immediately pipetted into a 60 ml sterile BD luer lock syringe. Samples were then hand filtered through 0.22 µm Sterivex-GP Pressure Filter Units and capped with B.Braun luer lock caps. 740 µl Buffer ATL and 60 µl Proteinase K from the Qiagen DNeasy Blood and Tissue Kit were added to each sample and they were placed in 50 ml falcon tubes in a rolling incubator overnight at 56°C. The solutions of Buffer ATL, Proteinase K and eDNA were transferred from the Sterivex-GP Pressure Filter Units to 2 ml microcentrifuge tubes using 10 ml BD Slip Tip Sterile Syringes (Fisher Scientific). DNA was then isolated using a modified Qiagen DNeasy Blood and Tissue Kit protocol.
samp_size
10 ml sand
type
false crawl nest surface
BioSampleModel
Metagenome or environmental