highly multiplex indexes for Hiseq

jiankuihe · 01-23-2013, 10:59 PM

Hello,

We plan to pool 30 samples into one lane. however, the TruSeq has only 12 indexes, and we are not going to buy Nextraseq kit. Is there any published barcode indexes sequences? We only need the index sequences

Thanks,
JK

kcchan · 01-24-2013, 10:20 AM

There are only a total of 24 unique indices available for the TruSeq LT kit. A TruSeq DNA HT kit includes 96 indices using a dual indexing method (12x8). That should give you enough barcodes to pool all of your samples. I wouldn't recommend making your own TruSeq adapters as they are a bit tricky.

kwaraska · 01-24-2013, 10:29 AM

Not only are they tricky to make-they are very expensive. Since the adapters are so long, they can't be made as standard oligos. They need a different process and purification to ensure they don't cross contaminate one another.

We are a core so it was worth it to us but we spent over $6k on 24 indexes.

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01-23-2013, 10:59 PM	#1
jiankuihe Junior Member Location: California Join Date: Dec 2011 Posts: 3	highly multiplex indexes for Hiseq Hello, We plan to pool 30 samples into one lane. however, the TruSeq has only 12 indexes, and we are not going to buy Nextraseq kit. Is there any published barcode indexes sequences? We only need the index sequences Thanks, JK

01-24-2013, 10:20 AM	#2
kcchan Senior Member Location: USA Join Date: Jul 2012 Posts: 185	There are only a total of 24 unique indices available for the TruSeq LT kit. A TruSeq DNA HT kit includes 96 indices using a dual indexing method (12x8). That should give you enough barcodes to pool all of your samples. I wouldn't recommend making your own TruSeq adapters as they are a bit tricky.

01-24-2013, 10:29 AM	#3
kwaraska Senior Member Location: Boston,MA Join Date: Nov 2008 Posts: 122	Not only are they tricky to make-they are very expensive. Since the adapters are so long, they can't be made as standard oligos. They need a different process and purification to ensure they don't cross contaminate one another. We are a core so it was worth it to us but we spent over $6k on 24 indexes.