Hi everyone,
We're currently attempting to make Illumina sequencing libraries with Epicentre's Ribo-Zero. Before sequencing, we TOPO-cloned our libraries and sequenced to clones (~20 per library) to get an idea of our libraries' diversity. Much to our dismay, we have found that a large portion of our clones (60-70%) map back to mitochondrial DNA. The predominant sequence bears a great deal of homology to the human gene Mt-rnr2-l (our libraries are from mouse). Has anyone else had similar difficulties with mitochondrial RNA? I've seen a few posts on this site where a higher mitochondrial RNA quantity has been referenced, but none addressing this problem specifically. I appreciate any help you can offer. Thanks in advance.
We're currently attempting to make Illumina sequencing libraries with Epicentre's Ribo-Zero. Before sequencing, we TOPO-cloned our libraries and sequenced to clones (~20 per library) to get an idea of our libraries' diversity. Much to our dismay, we have found that a large portion of our clones (60-70%) map back to mitochondrial DNA. The predominant sequence bears a great deal of homology to the human gene Mt-rnr2-l (our libraries are from mouse). Has anyone else had similar difficulties with mitochondrial RNA? I've seen a few posts on this site where a higher mitochondrial RNA quantity has been referenced, but none addressing this problem specifically. I appreciate any help you can offer. Thanks in advance.