QuantSeq kits enable cost-efficient sequencing for gene expression studies targeting 3' end of RNA transcripts. These kits are an exceptional alternative to standard RNA-Seq and microarrays.
We offer different expression profilling kits:
The QuantSeq FWD kit is designed to generate Illumina-compatible libraries of sequences close to the 3’ end of polyadenylated RNA
Analysis of Low Input and Low Quality Samples
QuantSeq kits are compatible with low input samples, down to 1 ng and are suited to work with low-quality input material, including FFPE samples.
The QuantSeq kit with UDI enables to get confident data output regarding gene counts and correlations from FFPE preparations or Fresh-Frozen material.
Mapping of Transcript End Sites (TES)
By using longer reads, QuantSeq FWD allows to exactly pinpoint the 3’ end of poly-A RNA and therefore obtain accurate information about the 3’ UTR.
High Strand Specificity
QuantSeq maintains exceptional high strand-specificity (>99.9%) and allows to map reads to their corresponding strand on the genome, enabling the discovery and quantification of antisense transcripts and overlapping genes.
Unique Molecular Identifiers (UMI)
The separately available UMI Second Strand Synthesis Module for QuantSeq FWD allows unique tagging of individual transcripts with 6nt long UMI’s, located between the partial P5 adapter and the random priming sequence.
This enables identifying PCR duplicates and eliminating amplification bias.
Direct Counting for Gene Expression Quantification
QuantSeq kits produce only 1 fragment per transcript. Therefore, no length normalization is required, allowing more accurate determination of gene expression values. This makes QuantSeq the best alternative to microarrays and conventional RNA-seq in Gene Expression studies.
In only 4.5 hours, including less then 2 hours hands-on time, QuantSeq’s straightforward workflow generates sequencing-ready NGS libraries
Cost Saving Multiplexing
The length restriction in QuantSeq libraries allows a high level of multiplexing by saving sequencing space and enables up to 9.216 libraries being sequenced in a single Illumina lane.
This high level of multiplexing allows saving costs as the length restriction in QuantSeq saves sequencing space.
Red: V1, Blue: V2, Green: V1 + 6 cycles, Violet: V2 + 6 cycles
When working with low input samples, overcycling may happen (non-counting errors). Avoid short libraries and increased amounts of amplification artefacts (such as linker-linker amplicons).
Do not worry about lost reads or even misassigned reads! UDI kits can adapt to most of commercially available library generation solutions.
The QuantSeq REV kit is designed to generate libraries at the 3’ end of poly-A RNA. This makes it possible to exactly pinpoint the 3’ end in Read 1 and enables the study of the 3’ UTR and alternative polyadenylation.
Depletion of globin mRNAs during QuantSeq Library Prep
Combined with the Globin Block Module, it is possible to generate globin-depleted, sequencing ready 3’ mRNA libraries from as little as 50ng of total RNA from whole blood.
The Globin Block module can be integrated seamlessly in the protocol without additional protocol steps. Morevover, it enables to reduce reads mapping to globin mRNA from 50-80% to as low as 5%. Consequently, the results enable a dramatically increase on gene detection.
Increased gene detection in human blood Quantseq libraries using Globin Block. CPM = Counts per million*
*: The input RNA was extracted using different methodologies = 1. SPLIT RNA Extraction kit 2. Other kit including red blood cell lysis or 3. Other kit without red blood cell lysis.
While maintaining exceptional strand-specificity of >99,9%, QuantSeq allows to map reads to their corresponding strand on the genome. This enables the discovery and quantification of antisense transcripts and overlapping genes.
Direct Counting for Gene Expression Quantification
Since only 1 fragment per transcript is produced, QuantSeq allows more accurate determination of gene expression values. Therefore, QuantSeq kits are the best alternative to microarrays and conventional RNA-seq in Gene Expression and eQTL studies.
Mapping of Transcript End sites
QuantSeq allows to exactly pinpoint the 3’ end of poly(A) RNA and therefore obtain accurate information about the 3’ UTR.
QuantSeq’s simple workflow allows generating ready-to sequence NGS libraries within only 4.5 hours, including less than 2 hours hands-on time.
The QuantSeq-Flex kit is a library preparation kit, designed to make Illumina compatible liraries from any RNA sample, using custom primers. Whether it is gene expression analysis, targeted sequencing, adapter-ligation based RNA-seq or any other experiment where a certain region needs to be sequenced, QuantSeq-Flex can be used together with user-supplied primers.
The following types of libraries can be generated:
The QuantSeq-Pool is the best solution for gene expression profiling for large screening projetcs using sample barcoding, early pooling and batch processing of up to 96 samples in one reaction. The workflow is easy scalable for multiplexing up to 36.864 samples.