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Old 10-15-2020, 09:41 AM   #1
Amygdala
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Location: United Kingdom

Join Date: Sep 2019
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Default Diluting Nextera XT Indexes?

Hey all,

My samples originate from LCM, and I am currently following the SMART-seq2 workflow. Recently one group has combined both these approaches and written a nice protocol for it: https://doi.org/10.1007/978-1-4939-7213-5_6

Both Picelli's SMART-seq2 protocol (for single cells) and the above LCM-seq protocols mentions diluting the index adapters from the Nextera XT Index Kit, which appears to go against Illumina's protocol for adding 5ul of each adapter per sample without diluting. In the Picelli's original paper the authors do not dilute the index adapters, so I am a bit confused.

Can anyone clear this up for me? Or explain the benefit of diluting the adapters? I would have thought that since I am adding the recommended 1ng of input material for the Nextera XT library prep kit that I would follow Illumina's protocol of not diluting the adapters.

Many thanks!
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