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Myoblast cell populations were isolated from muscle biopsies using the explant culture technique [23].
After homogenization, total RNA was isolated from the skeletal muscle biopsies using Trizol reagent from Invitrogen as specified by the manufacturer.
Total RNA was isolated from muscle biopsies using the RNeasy Kit (Qiagen) following the manufacturer's instructions.
Total RNA was extracted from muscle biopsies using a kit (RNeasy from Quiagen, Valencia, CA, USA), following the supplier's instructions.
Briefly, total RNA was extracted from muscle biopsies using TRIzol (Invitrogen Life Technologies Carlsbad, CA) according to manufacturer's protocol.
Genomic DNA was isolated from either primary cell lines or from muscle biopsies, using the DNeasy® Blood and Tissue kit (Qiagen, Valencia, CA, USA).
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Because of the limiting RNA amount isolated from muscle biopsies used for microarray analysis, only a few samples (6 cases) were used for confirmation with quantitative RT-PCR experiments.
Second, the muscle was biopsied using a Bergstroem muscle biopsy needle (Stille, Stockholm, Sweden).
The same five muscle biopsies were used for gene expression analyses by real-time reverse transcriptase polymerase chain reaction (RT-PCR) as described below.
Thus our conceptual framework centred on the muscle spindle and John, who had access to human intercostal muscle biopsies, subsequently used the same experimental rig to examine the muscle spindles in these muscles, and provided the first account of their activation in his in vitro preparation (Tom Sears; Supplementary material B8 ).
Histological and histochemical analyses were performed on 10 μm transversely orientated serial cryosections of skeletal muscle biopsy samples using standard procedures.
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