How to enter Unique Dual Index Primer into BaseSpace

gutiej · 08-16-2019, 02:07 PM

I'm using unique dual index primer pairs for my experiments. I'm multiplexing around 12 samples per NextSeq run. I don't know how to create a custom library in BaseSpace that will allow me to assign the unique primer pairs to all my samples as it's limited by a 12 x 8 format (12- i7 indexes and 8-i5 indexes, I need to be able to enter 12 i5 indexes). Any help would be greatly appreciated.

itstrieu · 08-16-2019, 02:35 PM

I ran into a similar issue recently. If you have the Dx version of the NextSeq, there is a software update that can do FASTQ generation on the machine similar to the MiSeq.

The other option is a work around where you will have to create two custom library kit. Each kit can do 8 UDI samples if you set your samples in a diagonal so that each row and column will have an unique index. After that just pool them together in the pool tab in BaseSpace.

gutiej · 08-16-2019, 04:08 PM

I don't have the Dx version but your workaround will be just fine, I didn't think it would let me combine samples from two different "kits". Thanks for the help!

RickC7 · 08-26-2019, 12:02 PM

https://support.illumina.com/bulleti...sequence-.html

Here's the bulletin from Illumina describing how to create a custom library prep kit in BaseSpace.

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08-16-2019, 02:07 PM	#1
gutiej Junior Member Location: California Join Date: Aug 2019 Posts: 2	How to enter Unique Dual Index Primer into BaseSpace I'm using unique dual index primer pairs for my experiments. I'm multiplexing around 12 samples per NextSeq run. I don't know how to create a custom library in BaseSpace that will allow me to assign the unique primer pairs to all my samples as it's limited by a 12 x 8 format (12- i7 indexes and 8-i5 indexes, I need to be able to enter 12 i5 indexes). Any help would be greatly appreciated.

08-26-2019, 12:02 PM	#4
RickC7 Member Location: Baton Rouge, Louisiana Join Date: Feb 2010 Posts: 31	custom library https://support.illumina.com/bulleti...sequence-.html Here's the bulletin from Illumina describing how to create a custom library prep kit in BaseSpace.

08-16-2019, 02:35 PM	#2
itstrieu Member Location: Atlanta Join Date: Nov 2018 Posts: 27	I ran into a similar issue recently. If you have the Dx version of the NextSeq, there is a software update that can do FASTQ generation on the machine similar to the MiSeq. The other option is a work around where you will have to create two custom library kit. Each kit can do 8 UDI samples if you set your samples in a diagonal so that each row and column will have an unique index. After that just pool them together in the pool tab in BaseSpace.

08-16-2019, 04:08 PM	#3
gutiej Junior Member Location: California Join Date: Aug 2019 Posts: 2	I don't have the Dx version but your workaround will be just fine, I didn't think it would let me combine samples from two different "kits". Thanks for the help!