Barcoded SMRTBell Adapters

[mzchr]

Hey everyone,

Another kind of barcoding question for pacbio (see fellow poster chammer's question on http://seqanswers.com/forums/showthread.php?t=44489 )
But we've also been thinking about multiplexing samples (>100-300) for Pacbio sequencing using barcoded SMRTbell adapters as we have limited input amounts of our DNA libraries. Also, these barcodes are (more or less) optimized for Pacbio, hence we'd favour these.

Could anyone share their experiences using this approach? Mainly in terms of advantages vs. disadvantages/limitations, etc.
For other approaches such as barcoded PCR primers you could theoretically multiplex 384 samples, whereas it seems that for barcoded adapters it seems to be only possible for 12 samples according to the following link;

http://www.smrtcommunity.com/servlet/servlet.FileDownload?file=00P7000000QKgshEAD

Could anyone clarify this for me? It'd be much appreciated!

Thanks!

m

[ECO]

I am interested as well. Not much info out there.

[rhall]

Not really my area of expertise, but I talked to someone who has worked with these barcodes:

There are a couple of issues with high multiplexing of barcoded adapters. One is that the library preps need to be done separately through ligation, which is most of the protocol. This is mentioned right at the beginning of the new shared protocol. Separate prepping of >100-300 samples would be painful unless they have a very automated process. Second, the current set of barcoded adapters is limited to 12 because they have 7-base barcodes, restricted to that size due to the HPLC purification size limit for oligos from IDT of 60 bases.