Transcriptome_profiling_of_mutant_embryos_from_the_Mouse_Genetics_project__DMDD_. Transcriptome_profiling_of_mutant_embryos_from_the_Mouse_Genetics_project__DMDD_

Total RNA was extracted from embryonic lethal homozygous gene knockout and sibling embryos from the Mouse Genetics Project (http://www.dmdd.org.uk/). Protocol: Total RNA was extracted and DNase treated. After fragmentation RNA was enriched for the 3’ ends by pull down using an anchored polyT oligo attached to magnetic beads. An RNA oligo comprising part of the Illumina adapter 2 was ligated to the 5’ end of the captured RNA and the RNA was eluted from the beads. Reverse transcription was primed with an anchored polyT oligo with part of Illumina adapter 1 at the 5’ end followed by 12 random bases, then an 8 base indexing tag, then CG and 14 T bases. An Illumina library with full adapter sequence was produced by 20 cycles of PCR. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/本数据集的样本源自小鼠遗传学项目（Mouse Genetics Project，http://www.dmdd.org.uk/）中的胚胎致死型纯合基因敲除胚胎及其同窝对照胚胎，总RNA（Total RNA）由此类样本中提取获得。实验流程：首先提取总RNA并经脱氧核糖核酸酶（DNase）处理。将RNA片段化后，通过与磁珠偶联的锚定聚胸苷寡核苷酸（anchored polyT oligo）进行下拉富集，以获取3'端RNA片段。将包含Illumina接头2（Illumina adapter 2）部分序列的RNA寡核苷酸连接至捕获RNA的5'端，随后从磁珠上洗脱目标RNA。以锚定聚胸苷寡核苷酸为引物进行逆转录：该引物的5'端携带来自Illumina接头1（Illumina adapter 1）的部分序列，其后依次连接12个随机碱基、8个碱基的索引标签、CG序列与14个胸苷碱基。通过20轮PCR扩增，构建得到带有完整接头序列的Illumina测序文库。本数据属于预发布数据集的一部分。若需了解威康信托桑格研究所（Wellcome Trust Sanger Institute）共享的预发布数据的规范使用方式（包括出版暂缓相关细则），请访问http://www.sanger.ac.uk/datasharing/