Long non-coding RNAs detected in Plasmodium falciparum malaria using high-resolution DNA tiling microarray technology.. Plasmodium falciparum 3D7

Survey of transcriptional activity across 22.6% of the P. falciparum strain 3D7 genome. 872 protein coding genes and 60 putative P. falciparum lncRNAs identified under developmental regulation during the parasite's pathogenic human blood stage. Overall design: Total RNA was isolated from synchronous in vitro parasite cultures at 18, 24, 30, and 36 hours post invasion (hpi) of red blood cells. 1 μg of total RNA was then subjected to poly-A selective amplification using Message Amp II (Ambion), substituting biased dNTP/NTP mixes (2A/T/U:1C/G). The resulting aRNA was labeled with Superscript II reverse transcriptase (Invitrogen), using random hexamers, and either Cy3- or Cy5-dUTPs (Amersham).

针对恶性疟原虫（Plasmodium falciparum）3D7菌株22.6%的基因组区域开展转录活性全景分析。本研究共鉴定得到872个蛋白编码基因及60个推定的恶性疟原虫长链非编码RNA（long non-coding RNAs, lncRNAs），上述转录本在该寄生虫致病的人体血液阶段均受发育调控。
实验设计：从同步化的体外寄生虫培养体系中分离总RNA，采集红细胞入侵后18、24、30及36小时（hours post invasion, hpi）的培养样本。取1 μg总RNA，采用Message Amp II（Ambion公司）进行poly-A选择性扩增，以偏倚性dNTP/NTP混合液（2A/T/U:1C/G）替代常规混合体系。所得反义RNA（antisense RNA, aRNA）采用随机六聚体引物，通过Superscript II反转录酶（Invitrogen公司）进行荧光标记，并分别使用Cy3-dUTP或Cy5-dUTP（Amersham公司）作为荧光标记底物。