Targeted Cloning of Large Pathways
DNAtrap™ technology provides custom cloning of biosynthetic gene clusters and long operons up to 150 kb in length.
DNAtrap™️ Biosynthetic Gene Cluster Cloning Service
DNAtrap™ technology provides custom cloning of biosynthetic gene clusters (BGCs) up to 150 kb in length. Varigen’s CRISPR/Cas9-based technology is significantly faster than competing techniques as we can deliver intact gene cluster clones from your sequenced microbe in 10 days. We have recently broadened our technology to include custom cloning from both fungal and bacterial targets.
Recombinant BGCs in 10 days!
Varigen has extensive experience cloning large, difficult DNA fragments, such as polyketide synthase (PKS), non-ribosomal peptide synthase (NRPS), and hybrid BGCs, as well as difficult to PCR fragments like virulence factor pathways. In these examples, unique restriction digest fragment patterns match the in silico simulated digest pattern, demonstrating the entire genomic fragment was correctly cloned into the BAC plasmid vector.
We have had 100% success using Varigen’s technology platform to clone specific DNA fragments up to 40kb into our destination vector. The insert DNA fragments were precision-excised from fungal genomic DNA using Varigen’s CRISPR/Cas9 technology. Importantly, our fungal strains are diverse wild-type isolates, not conditioned laboratory strains. Varigen’s capabilities and technology have exceeded our most ambitious expectations.
Dr. Philippe ProchassonAdvantages
- 150 kb gene cluster clones from virtually any microbe, bacterial or fungal
- Enables capture of hard-to-amplify sequences
- Delivered, verified clones. No library screening required
- IP protection for recombinant clone (and activities thereof)
- Customer retains all rights and ownership
Exact
Precision capture and cloning of intact pathways up to 150 kb
Convenience
Send us your microbial sample & sequence and our scientists do the rest!
Speed
10 day turn around
Cost
Cheaper than other methods (e.g. TAR or traditional BAC cloning)
Flexibility
Multiple BAC vector options and any sequenced genome
Confidentiality
Customer retains all rights and ownership
I had worked on cloning a pathway by conventional methods for over three months with little success. Varigen was able to perform the targeted cloning of an entire bacterial secretion pathway in less than 10 days! The amount of time, money, and frustration that your service saved our lab cannot be overstated. I look forward to doing business with you again.
Priscilla Barger
Example Targets
- Large secondary metabolism pathways, including repetitive polyketide and nonribosomal peptide natural products
- Genomic islands, virulence factors, proviruses, antibiotic biosynthetic pathways, etc
- Long operons, amplification resistant genes
What you provide
- Microbial cells
- Whole-genome sequence
- Biosynthetic gene cluster target of interest
What you receive
- Sequence-verified BAC clone of your region of interest
- Turn around time of 10 days!
Options
- Novel, inducible dual-promoter vector
- Customizable bacterial artificial chromosome (BAC) vector
CALL: 608.444.9518
Varigen Biosciences
505 S. Rosa Road
Suite 15
Madison, WI 53719
ABOUT VARIGEN
Varigen Biosciences Corporation is an NIH- and NSF-funded effort to harness naturally occurring microbial diversity to develop the next generation of natural products through genomics and metagenomics. Varigen's team is led by researchers with over 125 years of combined experience.