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QuantSeq-Flex Targeted RNA-Seq Library Prep Kit V2 for Illumina

  • Flexible kit for targeted sequencing and molecular barcoding
  • Identification of known and unknown fusion transcripts
  • Use of custom-designed gene-specific primers for first
    and/or second-strand synthesis
  • Cost-efficient and flexible alternative to pre-designed sequencing panels
  • Sequencing of up to 9,126 samples/lane

QuantSeq-Flex Targeted RNA-Seq Library Prep Kit V2

The QuantSeq-Flex Kit is a library preparation protocol designed to make Illumina-compatible libraries from any RNA sample using custom primers. It is open for development by advanced RNA-Seq users based on their custom needs.

The kit is based on the QuantSeq FWD Kit for Illumina (Cat. No. 015) strategy, with compatible reagents and Read 1 linker sequence introduced by the second strand synthesis primer. Hence the reads generated during the NGS run directly correspond to the RNA sequence.

QuantSeq Flex can be ideally combined with our QuantSeq with UDI V2 kits, allowing for optimal library multiplexing with Lexogen’s patented 12 nt Unique Dual Indices (Cat. No. 191 to 196).

QuantSeq-Flex Kit provides modules for Reverse Transcription (RT) and SSS, allowing users to choose their own primers for either one of these steps or both of them. This offers maximum flexibility in the choice of targets and input material.

With this highly flexible QuantSeq kit the following types of libraries can be generated:

  1. OligodT primed in reverse transcription, random primed in second strand synthesis (QuantSeq 3’ mRNA-Seq)
  2. OligodT primed in reverse transcription, target-specifically primed in second strand synthesis (targeted 3’ mRNA-Seq)
  3. Target-specifically primed in reverse transcription, random primed in second strand synthesis (targeted RNA-Seq, allows for identification of novel fusions)
  4. Target-specifically primed in reverse transcription, target-specifically primed in second strand synthesis (targeted RNA-Seq, known targets detectable only)

Whether it is gene expression analysis, targeted sequencing, adapter ligation-based RNA-Seq or any other experiment where a certain RNA region needs to be sequenced, QuantSeq-Flex can be used together with user-supplied primers.


autoQuantSeq 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.

autoQuantSeq allows for an automated 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

QuantSeq automation on other platforms may also be possible. Please contact our 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 reagent, 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.

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, but not hardware or software issues linked to the original liquid handling instrument supplier.

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