Mondala Tony S, Salomon Daniel R, Head Steven R
The Scripps Research Institute, La Jolla, CA, USA.
Methods Mol Biol. 2010;632:1-26. doi: 10.1007/978-1-60761-663-4_1.
In this chapter, we describe numerous methods to extract RNA, DNA, and protein from tissue, represented by kidney transplant biopsies, and from peripheral blood cells collected at various clinical sites. Gene expression profiling and SNP-based genome-wide association studies are done using various microarray platforms. In addition, protocols that enable simultaneous protein purification from these clinical samples, enable additional strategies for understanding of the molecular processes involved in organ transplantation, immunosuppressive drug regimens, and the elements determining allograft success and failure. Successfully establishing a multicenter clinical study was essential to meet our objectives for subject enrollment and transplant outcomes. This chapter focuses on our experience setting up and coordinating clinical sample collection from multiple transplant centers for the purpose of microarray analysis.
在本章中，我们描述了从以肾移植活检为代表的组织以及在不同临床地点采集的外周血细胞中提取RNA、DNA和蛋白质的多种方法。基因表达谱分析和基于单核苷酸多态性（SNP）的全基因组关联研究是使用各种微阵列平台进行的。此外，能够从这些临床样本中同时纯化蛋白质的方案，为理解器官移植、免疫抑制药物治疗方案以及决定同种异体移植成败的因素所涉及的分子过程提供了额外的策略。成功开展一项多中心临床研究对于实现我们的受试者招募目标和移植结果至关重要。本章重点介绍我们为进行微阵列分析而建立和协调从多个移植中心采集临床样本的经验。
