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HIGH SCHOOL

Biology

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Molecular Biology

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Studying DNA and RNA

Southern Blot for DNA Detection

Description

Southern blotting is a DNA detection method used after agarose gel electrophoresis separates DNA fragments by size. A DNA ladder with fragments of known length is run beside the sample so the fragment size can be estimated. The blot adds another step when scientists need to confirm the sequence identity of the DNA.

Transcript

Southern印迹是一种利用标记的DNA探针与目标DNA杂交,以检测基因组中特定序列的技术。

该技术以英国生物学家埃德温·南(Edwin Southern)的名字命名,他首次开发了这一技术。

该实验流程包括五个主要步骤:电泳、变性、膜转移、杂交和显影。

首先,基因组DNA经限制性内切酶消化,产生的DNA片段在琼脂糖凝胶上通过凝胶电泳进行分离。

为了使双链DNA变性或分离成两条单链DNA(ssDNA),将凝胶浸泡在氢氧化钠溶液中。

然后将凝胶置于含有氯化钠和Tris缓冲液的DNA中和溶液中的海绵上,以将其pH值重置为7.0。

接下来,将尼龙膜置于凝胶上方,并用一叠纸巾压住。单链DNA通过毛细作用转移到膜...

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