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Approximately 0.5 mg of DNA from various treated mice was digested and analyzed for titanium content.
10 20 µg genomic DNA from the tails of mice was digested with the indicated restriction enzymes and analyzed by Southern blot using a-P32-dCTP labeled random-primed DNA probes (Stratagene Prime-It II kit).
To sequence the transgenic insertion at the Metrn locus, genomic DNA from tgε26+/+ mice was digested with NcoI, self-ligated, and amplified using inversely oriented PCR primers (Fig. 2H, i-1F, i-1R, i-2F, and i-2R).
For NAD(P H measurements from primary L cells, SI tissue from GLU-Venus mice was digested to single cells [ 7] and analysed on the same day.
Articular cartilage from forelimb and hind limb joints of 6 week, 20 week, and 10 month old control and DNIIR mice was digested for 3 hours in 3 mg/ml collagenase D (Roche Applied Science) dissolved in DMEM/F-12 nutrient mixture (Gibco/Life Technologies, Grand Island, NY, USA) at 37°C in a thermal incubator.
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Tumor tissues of G3- and vector-expression cell treated mice were digested and lysated.
To genotype mice, tail biopsies from 3 week old mice were digested overnight at 55°C in Direct PCR Tail lysis buffer (Qiagen) and proteinase K (Qiagen).
The proximal region of prostatic ducts (i.e., the portion of the ducts nearest the urethra) of 6 week old C57BL/6 mice were digested and cells were examined for antigen expression (Table S1) [2], [2].
Pancreata from NODscid or Balb/c mice were digested with 1 mg/ml Liberase (Sigma-Aldrich, St . Louis MO) for 11 minutes in 37°C and then purified islets were separated using Histopaque-1077 (Sigma-Aldrich, St . Louis MO) followed by hand picking.
For osteoblast cultures, calvariae of neonatal mice were digested by 0.1% collagenase and 0.2% dispase 5 times, and cells isolated by the last 3 digestions were combined and cultured in α-minimal essential medium (α-MEM) containing 10% FBS.
Livers from wild type and MMTV-Cre mice were digested in buffer containing 2 mg/ml protease K, 10 mM Tris pH 8.0, 25 mM EDTA, 0.5% SDS, 100 mM NaCl at 55°C overnight.
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