核酸
胚胎
生物
溶解
RNA提取
互补DNA
核糖核酸
DNA
DNA甲基化
细胞生物学
分子生物学
生物化学
基因表达
基因
作者
Sarah Rose Huffman,Md Almamun,Rocío Melissa Rivera
标识
DOI:10.1007/978-1-62703-011-3_13
摘要
Researchers whose experimental models are mammalian oocytes and preimplantation embryos are often limited by the yield of nucleic acids that can be isolated from such a small sample size. In addition, the limited number of cells from these types of samples makes the simultaneous recovery of RNA and DNA very difficult and often sample pooling is necessary to increase nucleic acid yield. Here we report a simple set of procedures using commercially available kits that results in consistent yield and quality of nucleic acids. After sample lysis, RNA is isolated and converted to a reusable cDNA library. Following RNA isolation, DNA is precipitated, isolated, and bisulfite converted for DNA methylation studies. Our results demonstrate the feasibility of isolating RNA and DNA from a small number of cells with repeatability of results.
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