期刊
NATURE PROTOCOLS
卷 4, 期 6, 页码 902-922出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2009.61
关键词
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资金
- NIH, National Cancer Institute, Center for Cancer Research
- EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT [ZIAHD008877] Funding Source: NIH RePORTER
- NATIONAL CANCER INSTITUTE [ZIABC011081, Z01BC011081, ZIASC010437, ZICBC011068] Funding Source: NIH RePORTER
Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is a valuable tool for measuring gene expression in biological samples. However, unique challenges are encountered when studies are performed on cells microdissected from tissues derived from animal models or the clinic, including specimen-related issues, variability of RNA template quality and quantity, and normalization. qRT-PCR using small amounts of mRNA derived from dissected cell populations requires adaptation of standard methods to allow meaningful comparisons across sample sets. The protocol described here presents the rationale, technical steps, normalization strategy and data analysis necessary to generate reliable gene expression measurements of transcripts from dissected samples. The entire protocol from tissue microdissection through qRT-PCR analysis requires similar to 16 h.
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