Gausson Valérie, Saleh Maria-Carla
CNRS URA 3015, Institut Pasteur, Viruses and RNAi group, Paris, France.
Methods Mol Biol. 2011;721:107-22. doi: 10.1007/978-1-61779-037-9_6.
At the current rate of technological progress, high-throughput sequencing of nucleic acids has become a commodity. These techniques are perfectly suitable for viral small RNAs sequencing and contribute to the understanding of many aspects of virus biology in the context of host-pathogen interaction. However, the generation of high quality data is still an issue and the preparation of small RNAs libraries that accurately reflect the viral siRNAs in the sample remains a challenge. In this chapter we describe how to clone and sequence libraries of viral small RNAs from infected insect samples (mosquito, drosophilidae, insect-derived cell lines).
按照当前的技术进步速度,核酸的高通量测序已成为一种常规手段。这些技术非常适合用于病毒小RNA测序,并有助于在宿主 - 病原体相互作用的背景下理解病毒生物学的许多方面。然而,生成高质量数据仍然是一个问题,制备能够准确反映样本中病毒小干扰RNA(siRNA)的小RNA文库仍然是一项挑战。在本章中,我们将描述如何从受感染的昆虫样本(蚊子、果蝇科、昆虫来源的细胞系)中克隆和测序病毒小RNA文库。