生命经纬知识库 >>所属分类 >> DNA技术   

southern blots-Molecular Biology

标签: 暂无标签

顶[0] 发表评论(10) 编辑词条

Hi can anyone answer this

I am having problems with my southern hybridization.

I am wondering if the DNA transfers to my membrane.

Is it possible that the DNA can wash into the solutions when denaturing and neutralizing?

 -alshia-

--------------------------------------------------------------------------------

Try running a control blot with plenty of DNA (salmon sperm or other) cut with random enzymes. View the nictrocellulose filter after every step if DNA can be seen-if it is, your blots are just fine as far as transfer issues are concerned.

Poor signals on a Southern are usually due to probe defects coupled with incorrect hybridization temperatures-rarely due to DNA washing away unless the filter is old, brittle or exposed to too much light in storage or to ammonia/HCl from a nearby chemicals closet. Useful results have also been achieved when you do the primer hybridization step without blocking and/or running a primer hyb at 25C. This way you will get a lot of noise with the signal and then noise can be cleared away by increasing temperature or other variations.

-phdconsult-

--------------------------------------------------------------------------------

附件列表


→如果您认为本词条还有待完善,请 编辑词条

上一篇Histochemical detection重组质体的检定 下一篇基因克隆

词条内容仅供参考,如果您需要解决具体问题
(尤其在法律、医学等领域),建议您咨询相关领域专业人士。
0

收藏到:  

词条信息

admin
admin
超级管理员
词条创建者 发短消息   

相关词条