Description

The QuantSeq-Flex Targeted RNA-Seq Kit V2 is an all-in-one library preparation protocol designed to make Illumina compatible libraries from any RNA sample using custom primers. It is based on the QuantSeq FWD kit and contains one or both of the QuantSeq-Flex Modules.

With the QuantSeq-Flex First Strand Synthesis Module the reverse transcription primer can be substituted with target-specific primer. Using the QuantSeq-Flex Second Strand Synthesis Module V2 custom primer can also be used for the second strand synthesis. Please note that the second strand synthesis reaction requires annealing temperatures of >45 °C. In case you need lower temperatures, please contact info@lexogen.com.

For multiplexing of up to 384 samples/lane, the Lexogen i5 6 nt Dual Indexing Add-on Kit (Cat. No. 047) can be used that allows the dual indexing with four additional i5 indices.

NOTE: As of February 2019 barcode plates have been renamed to Lexogen i7 6 nt Index Set (7001-7096). An evaluation tool to check the color balance of index subsets is available here.

The old version of QuantSeq-Flex (Cat. No. 033-035) is available on demand. If you wish to order it, please contact info@lexogen.com.

Features and Application:

  • Gene expression studies of custom targets
  • Targeted sequencing
  • Suitable for low quality RNA (including FFPE samples)
  • Molecular barcoding

Kit Contents:

  • Reagent Box
  • First Strand Synthesis Module and/or Second Strand Synthesis Module
  • Lexogen i7 6 nt Index Set (formerly named: i7 Index Plate) with i7 indices 7001-7024 or 7001-7096 depending on the kit size (24 or 96)
  • Purification Module with Magnetic Beads

System Compatibility:

  • HiSeq 2000/2500/3000/4000
  • NextSeq 500/550
  • MiSeq
  • MiniSeq
  • Genome Analyzer

Optional PCR Add-on Kit (96 rxn)

The PCR Add-on Kit is highly recommended since PCR cycle numbers depend on the input material (type, quality, and amount of RNA) and may differ from the values given in the User Guide. Using the PCR Add-on Kit over- or undercycling of libraries can be prevented. Please note that the user will need to provide SYBR Green I (or an equivalent fluorophore) for the qPCR assay.