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Full ORF Entry Clones (ORFeome Collaboration - OC)

Full ORF Entry Clones (ORFeome Collaboration - OC)

Formed in late 2005, the ORFeome Collaboration (OC) was established to provide the research community with an unrestricted source of fully sequence-validated human Full ORF clones, in a format allowing easy transfer of the ORF sequences into virtually any type of expression vector.
One of the main goals of the project is to provide at least one fully-sequenced Full ORF clone for each of the ~ 18500 currently defined human genes.

The ORFeome Collaboration primarily focuses on human genes; however, the following collections for other model organisms have also been developed:

  • Mouse Full ORF Entry Clones (OCAC)
  • Zebrafish Full ORF Entry Clones (OCAF)
  • Entry Clone Collections for Xenopus and Drosophila.

In addition to the Entry Clone Collections, researchers at Dana-Farber Cancer Institute and The Broad Institute have developed a collection of ready-to-use Human Full ORF Lentiviral Expression Clones (hORFeomeV8.1 CCSB-Broad), for transfection and study of protein expression in mammalian cells.  

  • Entry Clone collections from various species (human, mouse, zebrafish, Drosophila, Xenopus)
  • Ready-to-use human Expression Clone Collection
  • Fully-sequenced and annotated
  • Genome-scale collections, 73% of 20506 RefSeq human genes covered
  • Gateway-adapted ORFs allow for efficient and easy transfer into a variety of compatible prokaryotic, mammalian, viral, or insect expression vectors
  • Many ORFs available with or without stop codon (annotated in your search hit) enabling production of native proteins or C-terminal fusion constructs
  • High fidelity: Each CDS was amplified for only 25 cycles with gene-specific primers and KOD HiFi Polymerase (Novagen), greatly minimising the risk of PCR-induced mutations

Protocols / Clone Handling

Please note that the clone(s) are tested as negative in our phage contamination assay prior to shipping. However, these clones should still be handled with care as no phage assay can be guaranteed to be 100% accurate.

Clones should be stored at +4°C on receipt. As soon as possible, re-streak the clones onto the same medium and incubate overnight at 37°C to obtain single colonies. A selection of at least 10 single colonies should be picked and grown in LB broth containing the appropriate antibiotic +8 % glycerol for subsequent freezing at -70°C. These frozen stocks can then be re-grown for plasmid isolation and purification.

References


More information on the creation of these libraries can be found in the following publications:

The ORFeome Collaboration Nature Methods 13:191–192 (2016) doi:10.1038/nmeth.3776.

Lamesch, P., Li, N, Milstein, S et al. (2006) hORFeome v3.1: a resource of human open reading frames representing over 10,000 human genes. Genomics. 2007 Mar;89(3):307-15. Epub 2007 Jan 5. PMID: 17207965

Rual JF, Hirozane-Kishikawa T, Hao T, Bertin N, Li S, Dricot A, Li N, Rosenberg J, Lamesch P, Vidalain PO, Clingingsmith TR, Hartley JL, Esposito D, Cheo D, Moore T, Simmons B, Sequerra R, Bosak S, Doucette-Stamm L, Le Peuch C, Vandenhaute J, Cusick ME, Albala JS, Hill DE, Vidal M. Human ORFeome version 1.1: a platform for reverse proteomics. Genome Res 2004;14:2128-2135. PMCID: PMC528929


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