Millard Andrew D, Tiwari Bela
Department of Biological Sciences, University of Warwick, Coventry, UK.
Methods Mol Biol. 2009;502:193-226. doi: 10.1007/978-1-60327-565-1_12.
Gene expression microarrays offer the ability to monitor the expression of all phage genes over an infection cycle. However, there are relatively few reports to date of microarrays being used to investigate phage biology. This chapter aims to provide an overview of how to design and implement a microarray experiment to investigate phage biology. Given the nature of microarrays being specific to an organism, each will provide a number of unique issues. In this chapter, we outline the basic theory behind microarrays and provide details on how to implement a microarray experiment from the design of oligonucleotide probes through to the hybridisation of microarrays. The matter of designing oligonucleotide probes will be discussed with regards to how probe length, secondary structure, free energy, probe orientation and amplification all have to be taken into account. As means of an example, the conditions used for the hybridisation of an array designed to be specific to the cyanophage S-PM2 is detailed.
基因表达微阵列能够监测噬菌体在整个感染周期内所有基因的表达情况。然而，迄今为止，利用微阵列研究噬菌体生物学的报道相对较少。本章旨在概述如何设计和开展一项微阵列实验来研究噬菌体生物学。鉴于微阵列针对特定生物体的特性，每种微阵列都会带来一些独特的问题。在本章中，我们概述了微阵列背后的基本理论，并详细介绍了如何开展一项微阵列实验，从寡核苷酸探针的设计到微阵列的杂交。将讨论寡核苷酸探针的设计问题，涉及探针长度、二级结构、自由能、探针方向和扩增等方面都必须加以考虑。作为示例，详细介绍了用于与专门针对蓝藻噬菌体S-PM2设计的阵列进行杂交的条件。
