Transcription profiling of E12.5 mouse whole embryos and placentas pooled within 3 litters to validate 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 "dye-swapped" experimental design was used to assess the accuracy and precision of 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（E12.5）的小鼠完整胚胎与胎盘组织混合，提取总核糖核酸（RNA）。从上述RNA样本中制备经荧光标记且线性扩增的互补核糖核酸（cRNA）靶标，采用新型微阵列（microarray）设计开展杂交比对实验，以验证该系统的有效性。采用“染料交换”实验设计评估该系统的准确性与精密度。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探针特征位点，其设计初衷为检测NIA（美国国家衰老研究所）全套互补脱氧核糖核酸（cDNA）克隆集合所对应的转录本。