Description

The QuantSeq FWD Kit is an all-in-one library preparation protocol designed to generate Illumina compatible libraries of sequences close to the 3’ end of polyadenylated RNA.

The kit is ideal for gene expression counting as NGS reads are generated towards the poly(A) tail. To get the exact 3’ transcript end longer reads are necessary.

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. It is already an integrated part in the QuantSeq 3’mRNA-Seq FWD HT Kit for 384 preps.

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 FWD (Cat. No. 015.24, 015.96 and 015.2×96) is available on demand. If you wish to order it, please contact info@lexogen.com.

Features and Application:

  • Gene expression studies
  • Suitable for low quality RNA (including FFPE samples)
  • From as low as 100 pg total RNA

Kit Contents:

  • Reagent Box
  • Lexogen i7 6 nt Index Set (formerly named: i7 Index Plate) with i7 indices 7001-7024; 1x, 2x or 4x 7001-7096 depending on the kit size (24, 96, 192 or 384 preps)
  • Purification Module with Magnetic Beads
  • Only for QuantSeq FWD HT (Cat. No. 015.384): Lexogen i5 6 nt Dual Indexing Add-on Kit (5001-5004)

System Compatibility:

  • HiSeq 2000/2500/3000/4000
  • NextSeq 500/550 (only FWD)
  • 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.