凍熱細菌質體轉形法

Abstract

細菌轉形為分子生物學相關研究領域實驗室內操作最為普遍之實驗步驟。本文介紹我們所發展之一快速且簡易之細菌轉形法名為「凍熱細菌質體轉形法」，利用此一方法可快速且大量進行細菌之轉形，轉形效率可達8.7×10^6轉形株／μg質體DNA。此一凍熱處理細菌質體轉形法僅須15min，較現有大多數方法耗時75-90min可節省60-75min，顯著縮短轉形所需時間。 We report a freezing-warming method for transformation of E. coli. In this method, plasmid DNA were mixed with 100 μl ice cold competent cells, freezing treated in liquid nitrogen for 5 min followed with heat treatment at 37°C for 5min, then directly spread onto the selective agar plate pre-warmed to 37°C and cultured for 16-18 hr.