Hi everybody,
I was wondering whether it would be possible to treat a single cell with DNAse prior to reverse transcription. There are so many protocols out there that are doing it but they start from MICROGRAMS (or nanograms, at least) of RNA, but what about single cells? I guess most of the RNA would be degraded after incubation at 37 degrees and inactivation of the DNAse at 65 degrees.
My problem is that I would like to sequence the nuclear RNA with random primers and need to get rid of the genomic DNA.
For obvious reasons, it has to be a method that doesn´t imply any purification or transfer of solutions from one tube to another. Any suggestion is very much appreciated!
I was wondering whether it would be possible to treat a single cell with DNAse prior to reverse transcription. There are so many protocols out there that are doing it but they start from MICROGRAMS (or nanograms, at least) of RNA, but what about single cells? I guess most of the RNA would be degraded after incubation at 37 degrees and inactivation of the DNAse at 65 degrees.
My problem is that I would like to sequence the nuclear RNA with random primers and need to get rid of the genomic DNA.
For obvious reasons, it has to be a method that doesn´t imply any purification or transfer of solutions from one tube to another. Any suggestion is very much appreciated!