Paris Pamela L
Department of Urology, University of California at San Francisco, San Francisco, CA, USA.
Methods Mol Biol. 2009;556:89-98. doi: 10.1007/978-1-60327-192-9_7.
High-resolution genomic arrays and next-generation sequencers are some of the genome-based technologies poised to make significant contributions in the near future to basic and clinical science. The success of these technologies, and most certainly their translation into the clinic, will require that they produce high quality, reproducible data from small archived tumor specimens, including biopsies. DNA from patient samples, especially archival tissue, can be a limiting factor and lead to the need for amplification of the starting material. A variety of whole-genome amplification techniques are available, but choosing the most reliable, reproducible amplification technology that will be suitable for use across a wide spectrum of clinical specimens is essential. Sigma's whole-genome amplification kit provides a robust, highly reliable, and versatile amplification system across a variety of DNA sources. This chapter will detail Sigma's amplification protocol along with an optimized DNA extraction protocol for formalin-fixed and paraffin-embedded tissue.
高分辨率基因组芯片和下一代测序仪是一些基于基因组的技术,有望在不久的将来为基础科学和临床科学做出重大贡献。这些技术的成功,以及它们肯定会在临床中的应用,将要求它们从小的存档肿瘤标本(包括活检组织)中产生高质量、可重复的数据。来自患者样本的DNA,尤其是存档组织的DNA,可能是一个限制因素,并导致需要对起始材料进行扩增。有多种全基因组扩增技术可供选择,但选择最可靠、可重复的扩增技术,使其适用于广泛的临床标本至关重要。西格玛的全基因组扩增试剂盒提供了一个强大、高度可靠且通用的跨多种DNA来源的扩增系统。本章将详细介绍西格玛的扩增方案以及针对福尔马林固定石蜡包埋组织的优化DNA提取方案。