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Studying DNA and RNA
Purifying DNA Bands from Agarose Gels
Description
Agarose gel electrophoresis can be used to purify DNA bands after separation by size. In gel extraction, the DNA fragment of interest is cut from the gel and cleaned before later use. This is especially useful in cloning, where pure DNA is needed before ligation, the joining step that links DNA pieces together.
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Transcript
琼脂糖凝胶电泳是一种常用于根据DNA分子大小对其进行分离的技术。
琼脂糖——一种源自红藻的多糖——加入如Tris-乙酸盐EDTA(TAE)或Tris-硼酸盐EDTA(TBE)等缓冲液中,并通过加热溶解。
通常,将 1 克琼脂糖溶解在 100 毫升缓冲液中以形成 1% 的琼脂糖溶液,但根据具体应用的不同,可添加不同量的琼脂糖。
为了染色凝胶,通常将荧光染料(如溴化乙锭)加入琼脂糖溶液中。该染料可插入DNA碱基对之间,并在紫外光下发出荧光。
然后可将溶液倒入水平模具中。将梳子插入凝胶模具中,以形成加样孔,用于加载DNA样品。
琼脂糖溶液静置,使其凝固成凝胶。
凝胶由通过氢键结合的琼脂糖分子构成,形成多孔基质...
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