Author
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Chao, Wun |
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Submitted to: Austin Journal of Plant Biology
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 10/20/2014 Publication Date: 10/27/2014 Citation: Chao, W.S. 2014. Ethidium bromide-free DNA in chimeric gene construction involving multiple DNA segments. Austin Journal of Plant Biology. 1(1):2. Interpretive Summary: This manuscript describes a method of extracting DNA from agarose gels without exposure to ethidium bromide (EtBr) and UV radiation. Although it is well known that DNA exposed to EtBr and UV is less effective for cloning work, EtBr is still the most widely used staining agent today. Another, newer and well-liked staining agent, SYBR Green, also requires UV radiation. Here I describe a new and simple method that uses these high sensitive fluorescent agents to locate DNA for extraction, but the DNA itself is not exposed to either fluorescent dyes or UV. Thus, the purified DNA can be used effectively for multi-segment ligation and cloning work. Technical Abstract: This manuscript describes a method of extracting DNA from agarose gels without exposure to ethidium bromide (EtBr) and UV radiation. Although it is well known that DNA exposed to EtBr and UV is less effective for cloning work, EtBr is still the most widely used staining agent today. Another, newer and well-liked staining agent, SYBR Green, also requires UV radiation. Here I describe a new and simple method that uses these high sensitive fluorescent agents to locate DNA for extraction, but the DNA itself is not exposed to either fluorescent dyes or UV. Thus, the purified DNA can be used effectively for multi-segment ligation and cloning work. |
