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The updated kits are provided with a revised QuantSeq-Pool User Guide (139UG272V0300) , which that is already available online.
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Our Quality Control Monitoring program detected a reduction in library pool yields after 1 year of storage. We have verified carefully that this yield decrease does not impact library data quality. The cause was traced to the inclusion of DTT within the First Strand cDNA Synthesis Component Mix (FS). To ensure robust stability of FS over the guaranteed shelf-life of 12 months, we now provide DTT and FS as separate components an additional component, DTT that is combined with FS for First Strand cDNA Synthesis.
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Product Component changes
FS no longer contains DTT composition was adjusted for increased stability.
Additional component DTT is provided as a separate tube.
Protocol changes
In addition to the component changes, our team verified that elevated temperature handling during first strand cDNA synthesis was not critical for library yield or data analysis quality. Therefore, an additional protocol update was also included.
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Step 2: Reactions are cooled and to, then held at 25 °C instead of 42 °C after denaturing the RNA / RT Primer mix.
Step 3: The mastermix is prepared with FS + DTT + Enzyme Mix 1 (E1), . Prewarming of this mastermix at 42 °C is no longer required and was removed from the protocol.
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The final reaction volumes and composition of the first strand cDNA synthesis reaction do not change due to the separate addition of DTT.
The reduced holding temperature at Step 2 and omission of mastermix pre-warming has no negative impacts on library preparation or data quality.
There is no negative impact on data quality if libraries were already prepared with kits purchased prior to November 2021.
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