Transcription profiling of E12.5 mouse whole embryos and placentas pooled within 3 litters and used to valdiate a new array

E12.5 mouse whole embryos and placentas were pooled within 3 litters and total RNA was extracted. Fluorescently-labeled, linearly-amplified cRNA targets were prepared from the RNA samples and compared on a novel microarray design to validate the system. A "self-versus-self" experimental design was used to assess the false-positive rates in the system. The in situ-synthesized 60-mer oligonucleotide microarray platform contains approximately 22,000 DNA features, and was designed to detect transcripts from the entire NIA cDNA clone collection.将3窝胚胎发育第12.5天的小鼠完整胚胎与胎盘混合，提取总RNA。从上述RNA样本中制备荧光标记、线性扩增的互补核糖核酸（cRNA）靶标，采用新型微阵列芯片设计进行杂交比对，以验证该实验系统的有效性。本研究采用“自身-自身”对照实验设计，对该系统的假阳性率进行评估。本次使用的原位合成60聚体寡核苷酸微阵列平台包含约22000个DNA探针位点，其设计初衷为检测美国国家衰老研究所（National Institute on Aging, NIA）全套互补脱氧核糖核酸（cDNA）克隆集合对应的转录本。