I would like to know what the best DNA extraction method for NGS analysis generally we use phenol chloroform method for Cancerous tissue DNA isolation but i heard phenol inter pt the NGS analysis, so which manual method is used to isolate tissue DNA? my friend suggest to use guanidine hydro chloride method ,is it useful for my work ? What is the best procedure for tissue DNA isolation
It is the best method. Phenol should not be a problem after ethanol precipitation.
I used phenol chloroform extraction method for my metagenome sample, doesn't seem to has any problem at all.
Phenol/ Chloroform extraction is most robust and the yield is also very good but sometimes residual phenol may create some hindrance in any such downstream applications. For that it would be wise to use a kit based protocol like Qiagen kits. Because samples are always precious.
Thank you all
Mr. dhar is there any manual method to isolate DNA other than phenol chloroform method
Alkaline lysis method:
http://www.ncbi.nlm.nih.gov/pubmed/8214582
https://www.youtube.com/watch?v=WZbAfQ58eCw
I routinely use protease and guanidine treatments with downstream NGS. If you want an off the shelf kit and not make everything then Qiagen is in my opinion one of the better commercial kits. Remember that before beginning target prep you should have an idea of DNA quality such as functional copies and you can always test for presence of inhibitors in the isolated DNA by a qPCR approach.
hai jeffrey u routinely use protease and guanidine treatments with downstream NGS.
can u give brief protocol of that
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