Antoniou Eric, Taft Robert
The Jackson Laboratory, Bar Harbor, ME, USA.
Methods Mol Biol. 2012;825:237-51. doi: 10.1007/978-1-61779-436-0_18.
Second-generation sequencers such as the Illumina GAIIx make possible the study of all transcribed loci in a genome across an almost endless dynamic range. Although typical protocols call for starting from at least 1 μg of total RNA, this is not possible when studying small tissues or rare cell types. This chapter explains how to prepare Illumina sequencing libraries from mouse oocytes. The protocol is also suitable for mural and cumulus cells, flow sorted or laser captured cells.
第二代测序仪,如Illumina GAIIx,使得在几乎无限的动态范围内研究基因组中的所有转录位点成为可能。尽管典型的实验方案要求从至少1μg的总RNA开始,但在研究小组织或稀有细胞类型时这是不可能的。本章将解释如何从小鼠卵母细胞制备Illumina测序文库。该方案也适用于壁细胞和卵丘细胞、流式分选或激光捕获的细胞。