QuantSeq-Pool 3' mRNA-Seq Library Prep Kit - October 2022
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 (see Version: 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.
Protocol Changes
Due to the inclusion of the Removal Enzyme the following protocol steps were updated (additions are in italics):
First Strand cDNA Synthesis - Reverse Transcription
Step 4: Incubate for 15 minutes at 42 °C, then for 10 minutes at 70 °C and 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 for 15 minutes at 37 °C then 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.
These protocol modifications, and inclusion of the Removal Enzyme (RE) ensure consistent performance of the QuantSeq-Pool protocol across different sample types.
However, please note that the original QuantSeq-Pool protocol can still be used with any QuantSeq-Pool kits available for sale, simply by refering to the previous version of the QuantSeq-Pool User Guide (see Previous Version: 139UG272V0300).
Need further information?
If you have purchased a kit prior to October 24th 2022 and would like to use the updated protocol, or if you have any technical questions or feedback, please get in touch with our Technical Support team via support@lexogen.com.