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Hi,
I'm trying to get a conceptual understanding of how low biomass samples impact the number of sequencing reads that can be obtained during shotgun sequencing on an Illumina platform. If I have a low biomass sample such as from blood or air, and am targeting 10 Gbases my understanding is that I may end up with far fewer than 10 Gbases of sequencing data. Is this because all available DNA has been used and thus it is not possible to generate more DNA reads, or is this too simple a conceptual view? In such cases, is it reasonable to say that the DNA has been sequenced to exhaustion and thus you have a full picture of the microbial community present in the (low biomass) sample. This would be in contrast to a high biomass sample such as soil where despite deep sequencing one could pragmatically always sequence deeper and it is understood that we may not get a single read from rare organisms.
Thanks,
Donovan
I'm trying to get a conceptual understanding of how low biomass samples impact the number of sequencing reads that can be obtained during shotgun sequencing on an Illumina platform. If I have a low biomass sample such as from blood or air, and am targeting 10 Gbases my understanding is that I may end up with far fewer than 10 Gbases of sequencing data. Is this because all available DNA has been used and thus it is not possible to generate more DNA reads, or is this too simple a conceptual view? In such cases, is it reasonable to say that the DNA has been sequenced to exhaustion and thus you have a full picture of the microbial community present in the (low biomass) sample. This would be in contrast to a high biomass sample such as soil where despite deep sequencing one could pragmatically always sequence deeper and it is understood that we may not get a single read from rare organisms.
Thanks,
Donovan