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QuantSeq 3’ mRNA-Seq V2 Library Prep Kit with UDI
Get the utmost quality out of our well-established QuantSeq 3’ mRNA-Seq gene expression kits, thanks to the improved V2 series! With the increasing reading capacity of next-generation sequencers, Lexogen’s patented 12 nt-long Unique Dual Indices will help you preserve most of your precious reads.
If you already have a library preparation kit without UDI (e.g., QuantSeq-Flex, Reverse, or Forward), our UDI Add-on V2 modules are available for purchase separately. They are delivered together with an improved PCR system, which can be seamlessly adapted to previous QuantSeq kits. For QuantSeq-Pool kits, you only need the UDI Sets (without purchasing additional PCR enzyme and buffer).
Our UDI Add-on V2 kits are also compatible with library preparation kits from other vendors.
For more information, please reach out to our specialists at support@lexogen.com
Performance
Consistent correlation between fresh frozen and FFPE samples
Whether your samples come from FFPE preparations, or fresh frozen (FF) material, you can be confident that data output will provide consistent gene counts and correlations.
Figure 1 | Sequencing data obtained with 10 ng input from two different mouse spleen samples, 1M reads, data shown for 1 CPM, 5 CPM, and 10 CPM (from left to right).
RIN values: 3.7 (FF) and 2.9 (FFPE)
DV200 values: 90% (FF) and 9% (FFPE)
Excellent resistance against overcycling
Overcycling may happen when working with low input samples. This results in shorter libraries and increasing amounts of amplification artefacts (such as linker-linker amplicons). With our new, improved V2 kits, even 6 additional cycles will not affect the quality of your library.
Figure 2 | Agilent Bioanalyzer traces of four libraries amplified with QuantSeq V1 with UDI or V2 with UDI kits, 100 ng input. Red: V1, blue: V2, green: V1 + 6 cycles, violet: V2 + 6 cycles. The V2 technology prevents library shortening, even with substantial overcycling.
Optimal UDI deduplication capabilities
Lexogen’s patented 12 nt UDI allow for particularly high error correction rates, with optimally designed sequence-Levenshtein values. With Lexogen, you do not have to worry about lost reads or even misassigned reads! And you can benefit from our design even on libraries generated with other vendors’ kits since our UDI kits can adapt to most of the commercially available library generation solutions. For optimal demultiplexing and read rescue, we recommend Lexogen’s iDemux command line.
Please consult with us for more information: support@lexogen.com.
Figure 3 | Lexogen’s 12 nt design allows rescuing the majority of undetermined reads thanks to superior error correction features, thereby saving precious data (based on 96 pooled libraries; Illumina NextSeq500 run, demultiplexed with iDemux).
Workflow
Automation
Automated QuantSeq protocol for 3’ mRNA-Seq Library Preparation
Automating the process of library preparation has the advantage of avoiding sample tracking errors, dramatically increasing throughput, and saving hands-on time.
QuantSeq library preparation and has been successfully implemented on several liquid handlers:
- Perkin Elmer: Sciclone® / Zephyr®
- Hamilton: Microlab STAR / STARlet
- Agilent: NGS Workstation (NGS Bravo Option B)
- Beckman Coulter: Biomek FXP, Biomek i5, Biomek i7
- Eppendorf: EpMotion® 5075
- Opentrons® OT-2
- Tecan: DreamPrep® NGS
QuantSeq automation on other platforms may also be possible. Please contact support team for more information.
Two key parameters must always be considered when automating a protocol:
- volume optimization
- script compatibility
In some instances, our kits will provide enough reagents while, in other instances, you will need a higher reagent volume or script adjustments. At Lexogen, we will help you find the best solution, tailored to your needs.
Before starting any new project involving automation, please reach out to our experts at support@lexogen.com
Please, also consider liaising with the robotic platform support team – they will be able to share the most recent version of the script.
Lexogen gladly supports the implementation of Lexogen-manufactured kits on liquid handlers, but not hardware or software issues linked to the original liquid handling instrument supplier.
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FAQ
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Safety Data Sheet
If you need more information about our products, please contact us through support@lexogen.com or directly under +43 1 345 1212-41.
Ordering Information
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