Resequencing the same Illumina library several times

[sinnafoch]

Hi,

I hope I am in the right forum. I have a Illumina library made of total RNA for a metatranscriptomics study.
If I sequence the library several times under the same conditions, can I expect (more or less) the same results? Same number of reads? Same distribution of reads across different taxa?

many thanks,

Philipp

[nucacidhunter]

You can expect similar results, though you should expect some variation in read number around +/-10% which may affect quality of reads and hence results. For instance, if first sequencing run clustered in high side of recommended density and the subsequent run was clustered10% higher than first run that will affect quality adversely and consequently your analysis results. But if it clustered lower than first run you will get less reads possibly with higher quality and that is less likely to affect the results.