联合应用低熔点回收法与连接介导PCR法进行体内足迹研究

In Vivo Footprinting Study by Combined Low Melting Gel Recovery and Ligation Mediated PCR

连接介导PCR是一种单侧PCR技术，它的应用大大地促进了体内足迹等研究的进行。在应用这一方法时，我们最关心以下三个方面：敏感性，忠实性和引物设计的方便。初始连接介导的PCR方法对于引物的要求有时限制了此技术的应用，尤其是在研究AT含量较高的调控元件时。本文采用低熔点回收法，有效地去除了PCR反应后所剩余的第二引物，大大方便了第三引物的设计。

Ligation mediated PCR is a single-sided PCR technology, its application greatly facilitates the execution of in vivo footprinting study. Our main concern while using this method involves sensitivity, fidelity and the ease of primer design. The strict primer requirement in the original LM-PCR method sometimes restrict its application, especially for AT-rich regulatory elements. In this paper, a new method is introduced: after the exponential amplification and the PCR product was recovered by low melting point agarose gel amplification. The method can effectively remove the leftover second primer, and facilitates the design of third primer.