在用碱裂解法提取质粒的过程中除质粒以外的DNA和RNA是怎么除去的碱裂解法提取质粒的原理是什么
在用碱裂解法提取质粒的过程中除质粒以外的DNA和RNA是怎么除去的
碱裂解法提取质粒的原理是什么
RNA 对碱不稳定,在提取质粒时,RNA都降解成小片段,质粒电泳时跑在最前面的就是RNA条带。
Alkaline denatured bacterial chromosomal DNA and most of RNA,since they are much much bigger than plasmid DNA.Plasmid is still soluble and is in the solution under the same condition.Proteins also get denatured,also,chromosomal DNA is linked to the bacterial proteins,therefore they all are precipitated when you include the 3 M potassium acetate.
The principal is that alkaline denatures proteins,chromosomal DNA and most of RNA,but leave plasmid in the solution,then potassium acetate precipitates those denatured materials.Finally,alcohol (usually isopropanol ) is used to precipitate plasmid DNA.Plasmid DNA is obtained.