Hello,
Two doubts. I hope someone can give their opinion. I'm working with total RNA extracted from a sample that comes from a pig infected with our gram-negative bacteria. We are interested in RNA from bacteria. We have previously centrifugated in vivo sample to obtein only bacterial cells.
After this, we used Ribo-Zero gram negative bacteria kit to remove rRNA from total RNA extracted with phenol-clorophorm method.
1- I have a strong band after rRNA removal. See Bioanalyzer file attached. I would like to know if you recomend me to sequence this sample or do a fragment size selection (open to other alternatives). We are interested in total bacterial RNA and also in their sRNA.
2 - We still have not treated the RNA extracted sample with DNAse. When and how do you recomen to do it.
Thank you very much, Bernardo
Two doubts. I hope someone can give their opinion. I'm working with total RNA extracted from a sample that comes from a pig infected with our gram-negative bacteria. We are interested in RNA from bacteria. We have previously centrifugated in vivo sample to obtein only bacterial cells.
After this, we used Ribo-Zero gram negative bacteria kit to remove rRNA from total RNA extracted with phenol-clorophorm method.
1- I have a strong band after rRNA removal. See Bioanalyzer file attached. I would like to know if you recomend me to sequence this sample or do a fragment size selection (open to other alternatives). We are interested in total bacterial RNA and also in their sRNA.
2 - We still have not treated the RNA extracted sample with DNAse. When and how do you recomen to do it.
Thank you very much, Bernardo
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