, 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)
◊ UltraPure
TM Agarose (Invitrogen, Cat. # 15510-027)
◊ UltraPure
TM 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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Isolate and Analyze Total RNA from Cells Harvested by Laser Capture
Microdissection2.
Laser microdissection and nucleic acid purification - a Leica - QIAGEN
cooperation3.
Working With Laser Capture Microdissection Samples4.
Laser Capture Microdissection (LCM) for the Analysis of Macrophage Gene Expression from Atherosclerotic Lesions5.
Laser Capture Microdissection (LCM) and Expression Profiling of Long-Range Projection Neurons6.
Differential Gene Expression Analysis of Pure Cell Populations from Frozen UterineTissue Using Laser Capture Microdissection (LCM) and cDNA Microarrays7.
Laser Capture Microdissection of Cells Labeled with Enhanced Green Fluorescent Protein8.
Primary cells, rat heart muscle9.
High-throughput gene expression analysis to screen for anti-asthma drugs10.
Quantitative end-point RT-PCR gene
expression measurement using the
Agilent 2100 Bioanalyzer and
standardized RT-PCR11.
Cytosolic expression of Green Fluorescent Protein (GFP) and its derivatives
in the yeast Saccharomyces cerevisiae: Detection in vivo using
the Varian Cary Eclipse