FlexGen

FlexGen was a biotechnology company based in Leiden, Netherlands. FlexGen was a spin-off from Leiden University Medical Centre and Dutch Space (part of EADS) and had proprietary technologies for laser based in-situ synthesis of oligonucleotides and other biomolecules. On the 21st of December 2015, Flexgen Bv in Leiden (Zuid-Holland) was declared bankrupt by the court in Gelderland Source.

FlexGen B.V.
IndustryBiotechnology
Founded2004
Headquarters
Leiden
,
Netherlands
Area served
Worldwide
ProductsFlexArrayer, FleXelect custom oligopools
ServicesCustom oligopool synthesis
Websitehttp://www.flexgen.nl

Products

FleXelect

FleXelect oligopools consist of custom oligonucleotides in solution and can be used for in solution target enrichment prior to next generation DNA sequencing. Target enrichment or In solution hybrid selection is a method for genomic selection in an increasing number of applications[1][2] such as;

  1. Analysis of custom genomic regions of interest (e.g. specific genes, multiple variants and/or complete pathways).
  2. Analysis of Chromosomal translocation
  3. Validation of Single-nucleotide polymorphism or SNPs (typically after whole genome or whole exome studies)
  4. Other research and diagnostic applications (e.g. Synthetic biology)

An example of a recent application is testing of the BRCA1 and BRCA2 breast cancer risk genes[3]

FlexArrayer

The FlexArrayer is an in-house custom oligonucleotide synthesis instrument. The FlexArrayer facilitates high troughput synthesis of FleXelect oligopools for in-solution target enrichment as well as custom microarray production. The FlexArrayer is also applicable for array based re-sequencing.[4]

The FlexArrayer provides microarray and oligopool synthesis typically used by:

  1. Genomics centres and sequencing facilities
  2. Health and safety institutes & microbiology labs
  3. Technology innovators in the fields of: Surface chemistries, PNA's (Peptide nucleic acid), siRNA's (Small interfering RNA) and more

Technology

Production of microarrays and FleXelect oligopools is done with the FlexArrayer (see image) using proprietary technology. The FlexArrayer synthesizes custom probesets on a substrate based on oligonucleotide deprotection technology;

  1. Before the first oligonucleotide synthesis step the complete DNA microarray surface is covered by photolabile groups.
  2. Those spots the first nucleotide addition is to occur are individually activated by the laser .
  3. The nucleotide solution is washed over the microarray surface and the nucleotides chemically bind to the activated spots.
  4. All nucleotides contain a photolabile group that can in turn be activated. As many rounds of photoactivation and nucleotide addition are performed as are required to synthesize oligonucleotides of the desired length.
  5. This is repeated up to 60 times until the required sequences (up to 100.000) have been synthesized. Thus, the maximum length of any oligonucleotide produced on this platform is 60mer in length.
  6. The microarray is now ready to be used, alternatively the oligonucleotides can be cleaved off to produce FleXelect oligopools.
The FlexArrayer, an oligonucleotide synthesis instrument.

References
  1. Mamanova, L.; Coffey, A. J.; Scott, C. E.; Kozarewa, I.; Turner, E. H.; Kumar, A.; Howard, E.; Shendure, J.; Turner, D. J. (2010). "Target-enrichment strategies for next-generation sequencing". Nature Methods. 7 (2): 111–118. doi:10.1038/nmeth.1419. PMID 20111037.Mamanova, L.; Coffey, A. J.; Scott, C. E.; Kozarewa, I.; Turner, E. H.; Kumar, A.; Howard, E.; Shendure, J.; Turner, D. J. (2010). "Target-enrichment strategies for next-generation sequencing". Nature Methods. 7 (6): 479. doi:10.1038/nmeth0610-479c.
  2. Tewhey, R.; Nakano, M.; Wang, X.; Pabón-Peña, C.; Novak, B.; Giuffre, A.; Lin, E.; Happe, S.; Roberts, D. N.; Leproust, E. M.; Topol, E. J.; Harismendy, O.; Frazer, K. A. (2009). "Enrichment of sequencing targets from the human genome by solution hybridization". Genome Biology. 10 (10): R116. doi:10.1186/gb-2009-10-10-r116. PMC 2784331. PMID 19835619.
  3. Two European Labs Turn to Next-Gen Sequencing for BRCA1/2 Testing, INsequence, October 26, 2010.
  4. SM Carr et al., DNA "Re-Sequencing" with a microarray, 2008. Comp Biochem Physiol D, Genomics & Proteomics 3,1-11 (link)
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