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RNA from tumor biopsies was obtained using the TriReagent (Gibco BRL Grand Island, New York) RNA extraction kit following the manufacturer instructions.
Total RNA from patient biopsies was obtained using an RNeasy kit on a QiaCube apparatus (QIAGEN) and served as the template (250 ng) to generate cDNA (Superscript VILO cDNA Synthesis Kit; Invitrogen).
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Until the 1970s, biopsies were obtained using metal capsules attached to a suction device.
The biopsies were obtained using a Bergström biopsy needle after local anaesthesia in the ICU patients and just after sedation in the control patients.
Muscle biopsies were obtained using the percutaneous needle [55], [56] method using manual suction from the vastus lateralis under local anaesthetic (1% lidocaine) as described previously [57].
Skin biopsies were obtained using previously reported methods [ 14].
8-mm ear punch biopsies were obtained using STIEFEL biopsy punches (Schuco, Germany).
Esophageal biopsies were obtained using standard sized, Radial Jaw 4 biopsy forceps (Boston Scientific, Boston, MA, USA).
Therefore, we believe that >90% of biopsies were obtained using a stroke length of 33 mm.
Patient core biopsies were obtained using a 14-gauge spring-loaded gun, yielding 15-20 mg of tissue per biopsy.
Biopsies were obtained using a 7.5 mm internal diameter modified Bordier trephine using local anaesthetic infiltration and intravenous sedation.
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