Characterization of the piRNA Complex from Rat Testes

This data series contains the sequences and read counts of Piwi-interacting RNAs and other small RNAs from mouse and rat testes extracts. In a search for different classes of small RNAs that might be involved in transcriptional gene silencing, we encountered a novel class of small RNAs within mammalian testes. This study reports the identification of small RNAs and their relative abundance levels to each other based on the deep coverage of the cDNA library. Keywords: High-throughput pyrosequencing by the 454 Life Sciences Genome Sequencer 20 System We constructed a cDNA library from small RNAs purified from Q column fractions of testes extracts. Briefly, gel-purified small RNAs from the column fractions were ligated with adaptor oligos on both 3' and 5' ends of the small RNAs. Reverse-transcription and PCR amplification was then performed on the ligated small RNAs to yield a cDNA library. This cDNA library was finally converted by PCR into a single-stranded DNA library compatible for coupling to beads on the 454 Life Sciences Genome Sequencer 20 System. Reads and sequences of the small RNAs were processed from linker sequences, examined for matches to the mouse or rat genomes, and tallied.本数据集包含小鼠和大鼠睾丸提取物中的Piwi互作RNA（Piwi-interacting RNAs）及其他小RNA的序列与读长计数。为筛选可能参与转录基因沉默的各类小RNA，本研究在哺乳动物睾丸中发现了一类全新的小RNA。本研究基于互补DNA（cDNA）文库的深度测序覆盖度，报道了小RNA的鉴定结果及其相互间的相对丰度水平。 关键词：基于454生命科学公司基因组测序仪20系统的高通量焦磷酸测序 研究人员从睾丸提取物的Q柱层析组分中纯化得到小RNA，并以此构建cDNA文库。简言之，将柱层析组分中经凝胶纯化的小RNA，在其3'与5'两端分别连接接头寡核苷酸；随后对连接了接头的小RNA进行反转录与PCR扩增，得到cDNA文库。最终通过PCR将该cDNA文库转化为单链DNA文库，可适配454生命科学公司基因组测序仪20系统的磁珠偶联流程。研究人员对小RNA的读长与序列进行接头序列切除处理，比对小鼠或大鼠基因组并完成统计计数。