کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2549950 1124533 2006 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Optimization and validation of small quantity RNA profiling for identifying TNF responses in cultured human vascular endothelial cells
موضوعات مرتبط
علوم پزشکی و سلامت داروسازی، سم شناسی و علوم دارویی داروشناسی
پیش نمایش صفحه اول مقاله
Optimization and validation of small quantity RNA profiling for identifying TNF responses in cultured human vascular endothelial cells
چکیده انگلیسی

IntroductionAffymetrix oligonucleotide microarrays are widely used in basic and applied research (Lander, 1999 and Lockhart & Winzeler, 2000). The need for a significant amount of starting RNA has limited its use in applications where the amount of RNA is limiting, such as with Laser Captured Microdissection (LCM), small biopsies, or peripheral blood in rodent models. To overcome this limitation, various RNA amplification and labeling methods have been described, however, further optimization and validation of these methods are needed.MethodsHere we reported using the Arcturus technology to optimize amplification and labeling of small amounts of RNA for Affymetrix microarray studies. We assessed the technical feasibility and variation introduced by differences in starting RNA quantity and differences in technical performance by microarray hybridization.ResultsWe demonstrated that the current approach is reliable to amplify as little as 40 ng total RNA, and it is suitable for Affymetrix studies yielding satisfactory quantitative chip performance. We also showed that differences in labeling methods contribute more to variation than the differences in starting RNA quantity per se. As a model, we studied the well-documented TNF-induced inflammatory responses in cultured human vascular endothelial cells. We were able to recapitulate the TNF-induced responses using small RNA sample profiling.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmacological and Toxicological Methods - Volume 53, Issue 2, March–April 2006, Pages 152–159
نویسندگان
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