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Laser Capture Microdissection of muscle fiber populations and expression analysis by RT-PCR

, Cat. # 12-550-19)
◊ Acetylthiocholine Iodide (Sigma, Cat. # A-5751)
◊ Sodium Acetate (Sigma, Cat. # S-2889)
◊ Sodium Citrate (Fluka, Cat. # 71402)
◊ Cupric Sulfate (Sigma, Cat. # C-1297)
◊ CapSure HS LCM Caps (Arcturus, Cat. # LCM0214)
◊ FastStart Taq DNA Poymerase (Roche, Cat. # 2 032 926)
◊ dNTP Mix PCR Grade (Invitrogen, Cat. # 18427-013)
◊ UltraPureTM Agarose (Invitrogen, Cat. # 15510-027)
◊ UltraPureTM 10x TAE (Invitrogen, Cat. # 15558-026)
◊ Ethidium Bromide (Sigma, Cat. # E-1510)
◊ Tissue-Tek OCT Compound (Sakura, Cat. # 4538)
◊ DEPC Treated Water (Ambion, Cat. # 9920)


Equipment and Labware

◊ General equipment necessary:
◊ Disposable gloves
◊ Flow hood
◊ Dry ice
◊ Liquid nitrogen
◊ Ice or cold block
◊ Nuclease-free, aerosol resistant tips
◊ RNase-free microcentrifuge tubes
◊ Scale
◊ KimwipesTM or similar lint-free towels

The following laboratory equipment is required to microdissect muscle fiber properly:
◊ Cryostat
◊ Cryostat knife
◊ Arcturus PixCell II Laser Capture Microdissection System with fluorescence package
◊ QImaingTM Retiga 1300 cooled mono 12-bit digital camera

The following laboratory equipment is required for RNA isolation and analysis by RT-PCR:
◊ Incubation oven
◊ Microcentrifuge
◊ Arcturus alignment tray
◊ Thermo Cycler with heated lid

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