Big results from small samples: evaluation of amplification protocols for gene expression profiling.

@article{citeulike:1620604, abstract = {Microarrays have revolutionized many areas of biology due to our technical ability to quantify tens of thousands of transcripts within a single experiment. However, there are still many areas that cannot benefit from this technology due to the amount of biological material needed for microarray analysis. In response to this demand, chemistries have been developed that boast the capability of generating targets from nanogram amounts of total RnA, reflecting minimal amounts of biological material, on the order of several hundred or thousand cells. Herein, we describe the evaluation of four chemistries for RnA amplification in terms of reproducibility, sensitivity, accuracy, and comparability to results from a single round of T7 amplification. No evidence for false-positive measurements of differential expression was observed. In contrast, clear differences between chemistries in sensitivity and accuracy were detected. PCR validation showed an interaction of probe sequence on the array and target labeling chemistry, resulting in a chemistry-dependent probe set sensitivity varying over an order of magnitude.}, address = {Bionomics Research and Technology Center, University of Medicine and Dentistry of New Jersey, Piscataway, New Jersey 08857, USA.}, author = {Viale, A. and Li, J. and Tiesman, J. and Hester, S. and Massimi, A. and Griffin, C. and Grills, G. and Khitrov, G. and Lilley, K. and Knudtson, K. and Ward, B. and Kornacker, K. and Chu, C. Y. and Auer, H. and Brooks, A. I. }, citeulike-article-id = {1620604}, issn = {1524-0215}, journal = {J Biomol Tech}, month = {July}, number = {3}, pages = {150--161}, posted-at = {2008-03-05 06:57:11}, priority = {2}, title = {Big results from small samples: evaluation of amplification protocols for gene expression profiling.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/17595311}, volume = {18}, year = {2007} }

TY - JOUR ID - citeulike:1620604 L3 - citeulike-article-id:1620604 TI - Big results from small samples: evaluation of amplification protocols for gene expression profiling. JF - J Biomol Tech VL - 18 IS - 3 SP - 150 EP - 161 AD - Bionomics Research and Technology Center, University of Medicine and Dentistry of New Jersey, Piscataway, New Jersey 08857, USA. SN - 1524-0215 N2 - Microarrays have revolutionized many areas of biology due to our technical ability to quantify tens of thousands of transcripts within a single experiment. However, there are still many areas that cannot benefit from this technology due to the amount of biological material needed for microarray analysis. In response to this demand, chemistries have been developed that boast the capability of generating targets from nanogram amounts of total RnA, reflecting minimal amounts of biological material, on the order of several hundred or thousand cells. Herein, we describe the evaluation of four chemistries for RnA amplification in terms of reproducibility, sensitivity, accuracy, and comparability to results from a single round of T7 amplification. No evidence for false-positive measurements of differential expression was observed. In contrast, clear differences between chemistries in sensitivity and accuracy were detected. PCR validation showed an interaction of probe sequence on the array and target labeling chemistry, resulting in a chemistry-dependent probe set sensitivity varying over an order of magnitude. AU - Viale, A AU - Li, J AU - Tiesman, J AU - Hester, S AU - Massimi, A AU - Griffin, C AU - Grills, G AU - Khitrov, G AU - Lilley, K AU - Knudtson, K AU - Ward, B AU - Kornacker, K AU - Chu, CY AU - Auer, H AU - Brooks, AI PY - 2007/07// UR - http://view.ncbi.nlm.nih.gov/pubmed/17595311 ER -