Hello, I work for a company that regularly performs barcoded pyrosequencing assays. What type of questions do you have?

Yes, you can barcode pyrosequencing samples. Up to 10 barcodes will work without many bad quality output. I have done this routinely with 454 sequencing which is pyrosequencing but done using commercial machines. The concept should be same for traditional pyrosequencing too.

Depending on what you want to do you should be able to find all details in published material. If stated specifically we can help.

If you are interested in 16S molecular diversity check out http://researchandtesting.com/

Hi, Alessandra

We have been pyrosequencing 16S samples with 454 for quite a while and it works fine if you take care in some details. What do you need to know?

Thanks for your replies, we are planning the experiment at the moment, and I am in doubt if use MIDs as tags or other oligos which allow to evaluate more than 10 samples (Hamady et al 2008 NATURE methods), waht do you think about? thanks

Here is another method for generating your tagged and linkered amplicons in PCR http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2515157/?tool=pubmed

We use barcoded 16S 454 sequencing with up to 300 different samples per run. That gives us about ~3000-5000 reads per sample. We use the Golay barcodes, see e.g. http://www.ncbi.nlm.nih.gov/pubmed/20566857 (we use a different primer set but the same barcodes).