Quantitative RT-PCR gene expression analysis of laser microdissected tissue samples
نویسندگان
چکیده
منابع مشابه
One-step RT-PCR for screening microdissected tissue samples.
The technology of laser-capture microdissection (LCM) enables the easy extraction of single cells or defined groups of cells from a tissue section (1,2). This technique uses a laser pulse to activate a plastic polymer, which binds to and captures cells from a frozen or paraffin-embedded tissue section. Captured cells present on this solid support can then be used for nucleic acid and/or protein...
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This chapter describes a method for the rapid assessment of gene copy number in laser microdissected material using multiplex real-time polymerase chain reaction (PCR). Here a putative oncogene (ZNF217) was evaluated in a series of colon tumors, but the method is applicable to any locus for which a nucleic acid sequence is available. The preparation, laser microdissection, and optimum storage o...
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Laser capture microdissection (LCM)-enabled region-specific tissue analyses are critical to better understand complex multicellular processes. However, current proteomics workflows entail several manual sample preparation steps and are challenged by the microscopic mass-limited samples generated by LCM, impacting measurement robustness, quantification and throughput. Here, we coupled LCM with a...
متن کاملOne-step RT-PCR without initial RNA isolation sStep for laser-microdissected tissue sample.
One-step RT-PCR procedure without initial RNA extraction step is tested for laser microdissected tissue sample. Unfixed cryosections of liver and kidney tissue of male SD rats were cut using laser microdissection system and directly used as templates for RT-PCR study. To check the sensitivity, 5, 25, 125, and 625 hepatocytes were cut and put in PCR-tube. After DNase treatment and cDNA synthesis...
متن کاملNOTE Pathology One-Step RT-PCR without Initial RNA Isolation Step for Laser-Microdissected Tissue Sample
One-step RT-PCR procedure without initial RNA extraction step is tested for laser microdissected tissue sample. Unfixed cryosections of liver and kidney tissue of male SD rats were cut using laser microdissection system and directly used as templates for RTPCR study. To check the sensitivity, 5, 25, 125, and 625 hepatocytes were cut and put in PCR-tube. After DNase treatment and cDNA synthesis ...
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ژورنال
عنوان ژورنال: Nature Protocols
سال: 2009
ISSN: 1754-2189,1750-2799
DOI: 10.1038/nprot.2009.61