DNA contamination in mRNA seq prepare

[ccqqsunshine]

Dear all,

I am frustrated with the contamination of DNA in my RNA sample,while treated with DNAse I(takara.) and extracted with phenol again,it seams all RNA were lost.

Any reconmondations for inactivation the DNAse1 instead?
thans for all and best wishes.


chenchen

[shurjo]

Dear all,

I am frustrated with the contamination of DNA in my RNA sample,while treated with DNAse I(takara.) and extracted with phenol again,it seams all RNA were lost.

Any reconmondations for inactivation the DNAse1 instead?
thans for all and best wishes.


chenchen

I use the Qiagen RNase-Free DNase kit with the RNAeasy Mini columns and they work just fine for me. No inactivation needed, the DNase I is just washed out on the column.

[greigite]

We use Ambion's Turbo DNA-free and it works great. Using standard DNAse protocols resulted in significant fragmentation of our RNA, which maybe is what happened to the OP.

[CC_seqanswers]

Could you elaborate a bit on the experience with Turbo? It worked great for you in terms of very low gDNA reads in the data? good yield of RNA? Have you compared the data between treatment and non-treatment? Thanks!

CC

We use Ambion's Turbo DNA-free and it works great. Using standard DNAse protocols resulted in significant fragmentation of our RNA, which maybe is what happened to the OP.