Simple • Affordable • Fast
Next-Generation Sequencing | Powered by Click-Chemistry

Simplify Sequencing with ClickSeq™ Technology

Affordable, Efficient, and Cutting-Edge RNAseq and DNAseq Solutions

ClickSeq™ Library Prep is a unique method for RNAseq and DNAseq that uses ‘Click-Chemistry’ in place of commonly used biological reagents such as fragmentases and ligases. ClickSeq is a simple procedure and considerably cheaper than other commonly used alternatives.

Next-Generation Sequencing Kits:

In partnership with baseclick GmbH (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.

Advantages of the ClickSeq™ Platform:

- 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

Submit Request here

ClickSeq™ Library Prep

RNAseq without the fragmentation or ligation.

A schematic representation of the core ClickSeq approach is presented. At top, an RNA molecule is depicted with two random primers with p7 overhangs are being extended into cDNA molecules with 3 prime azido terminating groups. Below, 'Click Ligation' is illustrated taking the two azido-terminated cDNA molecules and the chemical ligation using Copper Catalysed Azide-Alkyne Cycloaddition (CuAAC) to form single-stranded cDNA molecules with p5 and p7 adaptors on either end. Finally, these adaptor-ligated molecules are amplified by PCR to yield a double-stranded DNA library.

ClickSeq Schematic

ClickSeq: Fragmentation-Free Next-Generation Sequencing via Click Ligation of Adaptors to Stochastically Terminated 3′-Azido cDNAs:

Routh et al. JMB 2015

ClickSeq was originally published in 2015 in the Journal of Molecular Biology as a method to make RNAseq libraries that do not contain artifactual chimeric reads to allow the detailed analysis of rare recombination events in RNA viruses.

Read the Paper

Poly(A)-ClickSeq™

3' end RNAseq for simplified gene expression and alternative polyadenylation analysis

A screen shot of a genome browser track view showing read coverage over human gene Akt1. The top track in orange depicts read coverage obtained using poly(A)-ClickSeq, with a clear peak of coverage reaching 1510 counts adjacent only to the 3 prime end of the annotated gene. Below, a track showing read coverage from random-primed ClickSeq shows read coverage across the body of the gene with notable peaks of coverage above the annotated exons of the Akt1 gene, but not the introns. At the very bottom, a BED track depicts the genetic structure of the Akt1 mRNA with exon-intron boundaries and untranslated regions indicated.

Read Coverage over Human Akt1 mRNA: Poly(A)-ClickSeq (PAC-Seq) vs RNAseq

Poly(A)-ClickSeq: click-chemistry for next-generation 3΄-end sequencing without RNA enrichment or fragmentation:

Routh et al. NAR 2017

A method for sequencing just the 3′ ends of eukaryotic messenger RNAs by priming from poly(A) tails and using AzATP, AzGTP and AzCTP to terminate RT-PCR just upstream of the poly(A) tail in the 3’ UTR was published in Nucleic Acids Research in 2017.

Read the Paper

Tiled-ClickSeq™

Targeted sequencing: Each tiled amplicon is derived from only one template-specific primer

A schematic illustration depictoing overlapping normally-distributed peaks of read coverage above and to the left of Tiled-primers indicated below each peak. Each primer is extended with dotted black lines indicating cDNA synthesis that has been terminated by an azido-nucleotide at the left 3 prime end of the cDNA. Below, a similar scheme is shown, expect the normally-distributed coverage peaks are much broader and the black cDNA lines are longer, demonstrating how the same Tiled primer sets can provide different read coverage over the target genome.

Tiled-ClickSeq™ Schematic

Tiled-ClickSeq for targeted sequencing of complete coronavirus genomes with simultaneous capture of RNA recombination and minority variants:

Jaworski et al. eLife 2021

Tiled-ClickSeq™ leverages the ClickSeq™ approach to perform complete genome or gene sequencing. Using multiple tiled primers, overlapping amplicons spanning the target are generated. Tiled-ClickSeq™ has been optimized for the whole genome sequencing of RNA viruses including SARS-CoV-2.

Read the Paper

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Simple • Affordable • Fast Next-Generation Sequencing | Powered by Click-Chemistry

Simplify Sequencing with ClickSeq™ Technology

Affordable, Efficient, and Cutting-Edge RNAseq and DNAseq Solutions

ClickSeq™ Library Prep is a unique method for RNAseq and DNAseq that uses ‘Click-Chemistry’ in place of commonly used biological reagents such as fragmentases and ligases. ClickSeq is a simple procedure and considerably cheaper than other commonly used alternatives.

Next-Generation Sequencing Kits:

In partnership with baseclick GmbH (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.

Advantages of the ClickSeq™ Platform:

Request our services or contact us: info@clickseq.com

Poly(A)-ClickSeq: click-chemistry for next-generation 3΄-end sequencing without RNA enrichment or fragmentation:

Tiled-ClickSeq for targeted sequencing of complete coronavirus genomes with simultaneous capture of RNA recombination and minority variants:

Stay Up To Date

Simple • Affordable • Fast
Next-Generation Sequencing | Powered by Click-Chemistry