Hi,
I am new to analysing next generation sequencing data but have recently got involved in a Solexa project that aims to detect differences in small RNAs between tumour and normal samples. I have the resulting fastq read files which have had the adapter removed.
What I would like to know is the best settings to use with maq map for this kind of analysis.
1) I need to have a cut-off on the size of the reads used. I think you would use something like "-1 15" to only look at reads above 15 length
2) I think it is normal to allow a mismatch of 2 ... is this ok for small RNA stuff?
3) Anything else I need to look at?
One thing I can't understand is that even when I had the mismatch setting set at 2 the number of mismatches for the best match for some of the reads were way above that even getting up to 6.
Lastly is there any good resources for MAQ and small RNAs?
Many Thanks
I am new to analysing next generation sequencing data but have recently got involved in a Solexa project that aims to detect differences in small RNAs between tumour and normal samples. I have the resulting fastq read files which have had the adapter removed.
What I would like to know is the best settings to use with maq map for this kind of analysis.
1) I need to have a cut-off on the size of the reads used. I think you would use something like "-1 15" to only look at reads above 15 length
2) I think it is normal to allow a mismatch of 2 ... is this ok for small RNA stuff?
3) Anything else I need to look at?
One thing I can't understand is that even when I had the mismatch setting set at 2 the number of mismatches for the best match for some of the reads were way above that even getting up to 6.
Lastly is there any good resources for MAQ and small RNAs?
Many Thanks
