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The release of the BSA protein from CFCA-Ap was done by using a shaking bath (SKAKER, Sk-300).
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Muscles were then digested in sterile filtered type 1 collagenase (0.2%) in DMEM using a shaking water bath for 2 hours at 37 degrees.
Fluids were maintained at 37°C using a shaking water bath.
The temperature of SBF was maintained by using a shaking water bath, and SBF was completely refreshed every day.
Hybridization of the single-stranded, biotin-labeled amplicons to membrane-bound probes on the strip followed by addition of conjugate, and substrate to detect visible band patterns on the strip was performed manually using a shaking water bath, Memmert-SV1422 (Memmert GmbH & CO.KG, Schwabach, Germany) at 45°C.
The residue was dried over night and then extracted with 250 ml water (H2O) by using a shaking water-bath at 70°C for 2 hours.
The resulting mixture was continuously shaken in a shaking bath with a speed of 200 shakes/min at 293 K for 3 h until equilibrium was reached.
Without delay the flasks were then transferred into a shaking bath controlled at 60°C.
Then, flasks were placed in a shaking bath at 37 °C and RbCl (800 μM) was added.
The incubation was carried out in a shaking bath overnight at 37°C.
The epidermis was separated and then digested with typsin (0.05%) and Dnase (0.1mg/ml) for 15 min in a shaking bath.
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