Hi, I'm a very very new beginner in sequencing field. I'm getting microbial gDNA from environment samples and am thinking of doing Illumina 16S rDNA sequencing (metagenomics and RNA-seq in the future). Currently there's nobody in my lab doing sequencing with complex microorganisms sample and my boss want me to prepare the library by myself, so I'm just thinking of asking some stupid questions here for help...
Currently I have 12 gDNA samples (ranging from ~10 ng to ~1000 ng) and just want to do a 16S sequencing test run to get some data. I followed the 16S protocol in Illumina's website and target at the V3-V4 region (~550 bp totally). In order to get an overlap in the final sequence data we have booked the MiSeq reagent V3 kit (I think with its 2x300 cycle this is the only kit that can provide us with an overlap) and want to perform the sequencing on a MiSeq. However it's said that v3 kit can generate 13.2-15 GB data and is used for hundreds of 16S samples, in this case I'm kind of wasting it with only 12 samples. Could I ask for some suggestions on this issue?
Also I'm thinking if I'm doing metagenomics in the future maybe in this case I can also perform a metagenomics sequencing together with the 16S sequencing, using this v3 kit. Since I'm now only doing a test run, I don't need the depth and coverage to be that much. Would you think this might be possible?
Thanks a million in advance!!!!!!
Currently I have 12 gDNA samples (ranging from ~10 ng to ~1000 ng) and just want to do a 16S sequencing test run to get some data. I followed the 16S protocol in Illumina's website and target at the V3-V4 region (~550 bp totally). In order to get an overlap in the final sequence data we have booked the MiSeq reagent V3 kit (I think with its 2x300 cycle this is the only kit that can provide us with an overlap) and want to perform the sequencing on a MiSeq. However it's said that v3 kit can generate 13.2-15 GB data and is used for hundreds of 16S samples, in this case I'm kind of wasting it with only 12 samples. Could I ask for some suggestions on this issue?
Also I'm thinking if I'm doing metagenomics in the future maybe in this case I can also perform a metagenomics sequencing together with the 16S sequencing, using this v3 kit. Since I'm now only doing a test run, I don't need the depth and coverage to be that much. Would you think this might be possible?
Thanks a million in advance!!!!!!
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