Question: Demultiplexing same barcodes for different lanes
gravatar for gessthomsxavier7
13 months ago by
gessthomsxavier720 wrote:

I'm trying to demultiplex an illumina Novaseq XP worflow S2 flowcell run with a samplesheet in which same barcodes are present for samples in different lanes an example



bcl2fastq -R /mnt/NovaNAS/NovaSeq/190428_A00452_0029_BHJWFFDMXX -p 56 -o . -l TRACE

as expected bcl2fastq gave barcode collision error

2019-04-30 06:07:30 [1360880] ERROR: bcl2fastq::common::Exception: 2019-Apr-30 06:07:30: Success (0): /TeamCityBuildAgent/work/556afd631a5b66d8/src/cxx/lib/layout/BarcodeCollisionDetector.cpp(187): Throw in function void bcl2fastq::layout::BarcodeCollisionDetector::handleCollision(const value_type&, const value_type&)
Dynamic exception type: boost::exception_detail::clone_impl<bcl2fastq::layout::BarcodeCollisionError>
std::exception::what: Identical barcode found for multiple samples in the sample sheet: ATTACTCG+TATAGCCT

Is there any way to demultiplex using this samplesheet or should I create seperate samplesheet and run twice.

ADD COMMENTlink modified 13 months ago by genomax83k • written 13 months ago by gessthomsxavier720

i have a same problem, do you fix it?

ADD REPLYlink written 12 months ago by ngocttb0
gravatar for Devon Ryan
13 months ago by
Devon Ryan95k
Freiburg, Germany
Devon Ryan95k wrote:

You appear to have put two samples on the same lane with the same barcode. I have no idea how you could expect that to work, but you'll have to delete one and accept that they'll be mixed together. That's unless the Lane1 and Lane2 aren't meant to be lanes, in which case they should be in the first column ("Lane") as 1 and 2. Your sheet is generally malformed and I'm surprised that bcl2fastq was able to salvage as much as it did. In general, don't make these sheets by hand unless you really know what you're doing, use IEM.

ADD COMMENTlink written 13 months ago by Devon Ryan95k
gravatar for genomax
13 months ago by
United States
genomax83k wrote:

illumina Novaseq XP worflow S2 flowcell run

This is the critical bit. It is an XP flowcell which allows one to have lanes. You just need to have your samplesheet contain these columns


If you wish to get a single file per sample then use --no-lane-splitting option.

ADD COMMENTlink modified 13 months ago • written 13 months ago by genomax83k
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