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 i5 Dual Indexing Add-on Kit for QuantSeq/SENSE (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: The QuantSeq protocol has been upgraded on 17.02.2017 and is now even more streamlined. The RS2 addition step is removed and the following steps (second strand synthesis and purifications) are adjusted. Barcodes have been renamed (i7 Index Plate (7001-7096), formerly Barcode Plate (BC01-96)) and rearranged for a better nucleotide balance for sequencing when only few samples are multiplexed. Moreover, all indices are now different from Illumina barcodes (former BC05 – TAATCG) has been removed. An evaluation tool to check the color balance of index subsets is available here.
Features & Benefits
- Gene expression studies
- Suitable for low quality RNA (including FFPE samples)
- From as low as 100 pg total RNA
- HiSeq 2000/2500/3000/4000
- NextSeq 500/550 (only FWD)
- Genome Analyzer
- Reagent Box
- 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)
- Barcode Plate with external i7 Barcode Sets 1-3, 1-12 or 2x 1-12 depending on the kit size (BC01-96)
- Purification Module with Magnetic Beads
- Only for 384 preps (QuantSeq FWD HT): i5 Dual Indexing Add-on Kit for QuantSeq/SENSE for Illumina (5001-5004)