I want to do virome analysis (including DNA and RNA viruses) from various tissues and organs. What would be the best approach? Which kit would be best for nucleic acid extraction and is there a way to amplify both types of viruses simultaneously?
You will want to filter your homogenate through a filter with a small pore size to selectively enrich for viruses and not the host cells (this will reduce the number of host derived reads). You then will need to make cDNA of the RNA viruses which you can then use for sequencing along with the DNA of the DNA viruses.
Robert Adolf Brinzer Thank you very much for your answer. Would you care to recommend a protocol for the procedure you mentioned?
Most of them are similar to this (Article Rapid Identification of Known and New RNA Viruses from Animal Tissues
). There are variances with how viral particles are pelleted such as PEG addition and ultracentrifugation. As for the cDNA synthesis you may want to use random hexamers.- Badges
- Science method