tRNA/5S rRNA depletion

[mateo]

Dear all,
I'd like to deplete tRNA and 5S rRNA from total RNA to clone and deep sequence primarily precursor miRNAs in eukaryotic cells. I'm assuming that pre-miRNA are at a very low concentration when compared to tRNA/5S so if these species are left in my sample preparation they will represent the vast majority of sequences. Any help and/or suggestions would greatly be appreciated.
Thanks.

[whw]

Gel purification can be a good way of eliminating larger species. I know that Bioo Scientific includes a gel purification step in their small RNA sequencing protocol.

[pmiguel]

Dear all,
I'd like to deplete tRNA and 5S rRNA from total RNA to clone and deep sequence primarily precursor miRNAs in eukaryotic cells. I'm assuming that pre-miRNA are at a very low concentration when compared to tRNA/5S so if these species are left in my sample preparation they will represent the vast majority of sequences. Any help and/or suggestions would greatly be appreciated.
Thanks.

Epicentre RiboZero Gold?

--
Phillip

[protist]

Epicentre RiboZero Gold?

--
Phillip

RiboZero Gold will not remove tRNA, its advantage is it will remove mitochondrial as well nuclear encoded rRNA. To my knowledge aside from size selection of below tRNA size (~70 nt) there is not an effective kit out there..

[mnkyboy]

You can see the tRNA pretty well in a gel. I would try the RZ beads to clean it up then gel purify.

Also it will probably be easier to purify out after library preparation than as RNA by gel.