Bacterial Diversity Identification With Normalized Libraries?

Question

[Marina Manrique]

Hi all,

I'd like to know if normalized libraries are being used in bacterial identification experiments based on 16s rRNA when you don't need to get the abundance of detected species but to get the maximum number of species present in your sample. For example to get a general picture of the species present in your sample and then, if you think it's worthy, sequence 16s amplicons to find out the relative abundance between the different species present in the sample.

The approach would be similar to that used when normalizing cDNAs libraries in RNA-seq experiments to get as many different transcripts as possible.

Do you know if this approach is being used?

Thanks,

marina

Answer 1

[Larry_Parnell]

I have never heard of normalized libraries being used in the manner you describe.

[Marina Manrique]

me neither, it's just an idea I had after working a lot with normalized libraries in transcriptomes and then moving to 16S analysis. I think it'd be interesting but I have no idea of the lab challenges it would have.

Answer 2

[Shellfishgene]

I'd say this only makes sense if you suspect that bacteria that are present in your sample in very low numbers have a significant impact on the questions you are asking.

Also, I wonder if the similarity of all the 16S sequences wouldn't make normalization difficult.