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ClickSeq™ and Poly(A)-ClickSeq™ kits now available!
Introducing our latest breakthroughs in Next-Generation Sequencing library preparation: the ClickSeq and Poly(A)-ClickSeq kits, in partnership with baseclick GmbH (baseclick.eu)
After rigorous development and validation, we are proud to present these cutting-edge solutions, designed to elevate your sequencing experiments to new heights of precision and efficiency. Whether you're pursuing random primed RNAseq or DNAseq analysis with ClickSeq, or focusing on 3' end transcriptomics using Poly(A)-ClickSeq, these innovative kits promise to revolutionize your sequencing workflow and empower you with unparalleled insights into the world of genomics.
Our 3 NGS kits are now available for purchase at baseclick.eu:
We have developed and validated the following ClickSeq-based kits for NGS library preparation:
ClickSeq: for random primed RNAseq or DNAseq
Poly(A)-ClickSeq: for 3' end focused transcriptomics
X-ClickSeq: for use with user-provided primers in the Reverse Transcription step, allowing for customized, targeted and tiled assays.
ClickSeq™ kit: fragmentation-free RNAseq
Applications:
Random-primed RNAseq approach
mRNA sequencing
Gene expression analysis
Splice variant, isoform analysis, and gene fusion discovery
RNA virus genomics and recombination analysis
Benefits
No fragmentation steps required
No enzymatic ligation steps, reduces artifactual recombination
Highly degraded and/or fragmented RNA can be processed
Stranded technique: Provides strand-of-origin information
Excellent for the detection of rare recombination events
Reduced sample input, as little as 10ng purified RNA required
Libraries generated in ~6 hours
Unique Molecular Identifiers (UMIs) available
Compatible with Illumina platforms
Visit baseclick.eu to purchase ClickSeq kits:
ClickSeq Schematic
RNA is reverse transcribed using a 6N-primer containing a partial Illumina p7 sequencing adapter. Reverse transcription is performed in the presence of azido-nucleotides that stochastically terminate cDNA synthesis.
cDNA is purified using SPRI magnetic beads.
Click-chemistry is used to chemically ligate the Illumina p5 sequencing adapter.
Click-ligated cDNA is purified using SPRI beads.
PCR fills the remainder of the i7 indexing adapter and amplifies the amount of dsDNA library.
A final bead purification and size selection yields sequencing-ready libraries.
Poly(A)-ClickSeq kit: 3' end RNAseq for gene expression and polyadenylation analysis
Applications:
Captures any polyadenylated RNAs
mRNA sequencing and quantification
Gene expression analysis
Poly(A)-site discovery
Alternative Polyadenylation (APA) Analysis
Benefits
No fragmentation steps required
No enrichment/depletion steps required. Removes potential biases and reduces cost, time and loss of samples
No enzymatic ligation steps, reduces artifactual recombination
Highly degraded and/or fragmented RNA can be processed
Stranded technique: Provides strand-of-origin information
Reduced sample input, as little as 100ng Total Cellular RNA required
Libraries generated in ~6 hours
Unique Molecular Identifiers (UMIs) available
10-20M reads per sample is sufficient for most applications
Compatible with Illumina platforms
Visit baseclick.eu to purchase Poly(A)-ClickSeq kits:
https://www.baseclick.eu/product/polya-clickseq-library-prep-kit/
Poly(A)-ClickSeq Schematic
Total cellular RNA is reverse transcribed using an oligo-dT(21) primer containing a partial Illumina p7 sequencing adapter. Reverse transcription is performed in the presence of azido nucleotides that stochastically terminate cDNA synthesis upstream of poly(A)-tail.
cDNA is purified using SPRI magnetic beads.
Click-chemistry is used to chemically ligate the Illumina p5 sequencing adapter.
Click-ligated cDNA is purified using SPRI beads.
PCR fills the remainder of the i7 indexing adapter and amplifies the amount of dsDNA library.
A final bead purification and size selection yields sequencing-ready libraries.
X-ClickSeq™ kit: customized, targeted NGS
Applications
X-ClickSeq kits provide the same reagents as the ClickSeq kits, except no primers are provided for the Reverse Transcription step. Rather, the user may use their own RT primer as per the requirements of their own assay. For example, a 9N random primer might be used instead of a 6N random primer; or a set of SARS-CoV-2 targeting tiled-primers might be used (see Jaworski et al. eLife, 2021). All other aspects of the library prep are otherwise identical to ClickSeq. The data analysis pipeline will also be altered as per the assay design.
Benefits
Multiple tiled primers may be used to targets of interest
No fragmentation steps required
No enzymatic ligation steps, reduces artifactual recombination
Highly degraded and/or fragmented RNA can be processed
Stranded technique: Provides strand-of-origin information
Excellent for the detection of rare recombination events
Libraries generated in ~6 hours
Unique Molecular Identifiers (UMIs) available
Compatible with Illumina platforms
Visit baseclick.eu to purchase X-ClickSeq kits:
https://www.baseclick.eu/product/x-clickseq-library-prep-kit/
X-ClickSeq Schematic
RNA (or DNA) from a sample is reverse transcribed using a user-provided primer that should contain a partial sequence of the Illumina p7 sequencing adapter. Reverse transcription is performed in the presence of azido-nucleotides that stochastically terminate cDNA synthesis.
cDNA is purified using SPRI magnetic beads.
Click-chemistry is used to chemically ligate the Illumina p5 sequencing adapter.
Click-ligated cDNA is purified using SPRI beads.
PCR fills the remainder of the i7 indexing adapter and amplifies the amount of dsDNA library.
A final bead purification and size selection yields sequencing-ready libraries.
Advantages of ClickSeq™:
No fragmentation required;
No enzymatic ligation;
Ultra-low artifactual chimera formation¹;
Specialized Transcriptomic applications;
Integrated analysis pipelines, from RNA to results;
Affordable library synthesis
RNA or DNA
Suitable for fragmented/degraded samples
Request our services or contact us: info@clickseq.com
