SAMN00738286
External Id
University of Toronto
INSDC center name
2011-10-12T00:00:00Z
INSDC first public
2018-06-12T15:48:23.449Z
INSDC last update
SRS266493
INSDC secondary accession
live
INSDC status
Plant
NCBI submission model
Plant.1.0
NCBI submission package
SRS266493
SRA accession
not provided
collection_date
USO-31
cultivar
Genomic DNA was purified from nuclei isolated from young leaves. Briefly, leaves were frozen in liquid nitrogen, extracted with homogenization buffer (10 mM Tris pH 9.5, 80 mM KCl, 10 mM EDTA, 1 mM spermidine, 1 mM spermine, 0.15% β-mercaptoethanol and 0.5% Triton X-100) with gentle stirring and the extract filtered through cheesecloth and Miracloth. The nuclei and cell debris were pelleted by centrifugation and resuspended in wash buffer (as above with 0.5% (v/v) Triton X-100 and 0.15% β-mercaptoethanol) using a paintbrush. After low-speed centrifugation to remove cell debris, the nuclei were recovered by centrifugation. The nuclei were resuspended in wash buffer and centrifuged three times to remove other organelles. Genomic DNA was isolated from purified nuclei by treatment with proteinase K, extraction with phenol-chloroform and precipitation with isopropanol. The precipitated DNA was spooled using a glass hook, washed with 70% ethanol and treated with RNase. The DNA was extracted with phenol-chloroform, precipitated with sodium acetate/ethanol, spooled with a glass hook, washed with 70% ethanol and air dried. The purified DNA was dissolved in TE buffer.
description
missing
geographic location (country and/or sea)
missing
geographic location (region and locality)
Cannabis sativa
http://purl.obolibrary.org/obo/NCBITaxon_3483
organism
young leaf
tissue
Cannabis sativa 'USO-31' genomic DNA
title