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Dear Customers,
Please note the following updates to the QuantSeq-Pool 3' mRNA-Seq Library Prep Kit and protocol (Cat. No. 139.96). These updates are applied directly to kits shipped as of . The kit pricing and Catalog Number (139.96) remain unchanged.
The revised User Guide (139UG272V0400), which outlines the updated protocol is already available online.
Product Component Changes
The Removal Enzyme (RE) was added to the kit for use at the RNA Removal stage of the protocol.
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Due to the inclusion of the Removal Enzyme the following protocol steps were updated (additions are in turquoise italics):
First Strand cDNA Synthesis - Reverse Transcription
Step 4:Incubate for 15 minutes at 42 °C, then for 10 minutes at 70 °Cand cool to 25 °C, proceed to pooling and purification.
RNA Removal
Step 15: Add 2 µl of Removal Solution (RS) and 1 µl of Removal Enzyme (RE) to each pool, mix well and spin down. OPTIONAL:when processing 3 or more pools we recommend preparing a mastermix of RS and RE and adding 3 µl of the RS / RE mastermix per pool.
Step 16: Incubate for15 minutes at 37 °Cthen for 10 minutes at 95 °C …(Added incubation step).
Second Strand Synthesis
Step 20: Incubate for 15 minutes at 30 °C…
Why was the kit updated?
At Lexogen, we continuously strive to ensure our products meet the evolving needs of our customers. Based on customer feedback, we realized that we could improve the performance consistency of QuantSeq-Pool with a few protocol modifications. The updated protocol has the added benefit of increased differential expression sensitivity.
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