摘要
介绍一种从琼脂糖凝胶同步回收DNA和琼脂糖的方法。利用0.25 mol/L异硫氰酸胍溶液(p H 8.0)溶解含有目的 DNA片段的的凝胶条,胶条溶解后,静置冰上10 min再加入预冷的异丙醇,琼脂糖呈颗粒状析出,通过离心即可初步分离DNA和琼脂糖。上清液用异丙醇沉淀回收DNA片段,利用50%PEG溶液沉淀琼脂糖。分别对0.2 kb、1 kb和10 kb长度的DNA片段进行回收,回收率分别为19.44%、36.40%、13.49%,回收的DNA纯度高,电泳条带清晰。琼脂糖均回收率为62.52%,回收琼脂糖脱水后的状态为白色颗粒。该方法切实可行,回收成本低廉,回收的DNA和琼脂糖可用于后续实验。
A method for recovery of DNA and agarose in synchrony from the agarose gel was introduced. Gel contained DNA was dissolved by 0.25 mol/L guanidinium thiocyanate（p H 8.0）. After the gel dissolving, the precooling isopropanol was added and put on ice for 10 min. The agarose was precipitated. DNA and agarose were separated by centrifugation. DNA fragments were precipitated with isopropanol. 50% PEG solution was added to precipitate the agarose. The recovery rate of 0.2 kb, 1.0 kb and 10 kb DNA fragments were19.44%, 36.40%, 13.49% respectively. The purity of recovered DNA fragments showed the clear electrophoresis bands. Agarose average recovery was 62.52%. The recovered agarose state was white particles.The results showed that our method was simple and low cost, and both the recovered DNA and agarose gel could be reused.
出处
《生命科学研究》
CAS
CSCD
2015年第4期299-302,共4页
Life Science Research
基金
深圳市科创委基金项目(JC201005280549A)
关键词
琼脂糖
DNA
同步回收
回收率
agarose
DNA
recovery in synchrony
recovery rate
