Daniokey Pilot BAC clones
The Daniokey Pilot BAC library was created by the Ronald Plasterk Laboratory and Keygene N.V. The library provides approximately 1x coverage of the Zebrafish genome.
The library was created from the Tübingen zebrafish strain male testis DNA, provided by R. Geisler. The DNA was partially digested with HindIII. The DNA was cloned into the pIndigoBAC-536 vector, in E.coli DH10B.
There are 11,808 clones with an average insert size of around 130kb.
Source BioScience offers the complete clone set of 31 384-well microtitre plates or individual clones, for research purposes only. We can also make subsets of your chosen clones from the collection. Please contact us to discuss your requirements.
To examine the genomic organization of genes in zebrafish.
Order Conditions
The use of the resource is limted to research puproses. The collection is subject to the following MTA and our clone terms and conditions.
Due to MTA requirements, Daniokey Pilot BAC clones should be ordered online using our website search below, using the clone ID format e.g.DKEYp-118E10 or zkp118E10. to search for individual clones.
The entire library can be ordered below:
| Product Code |
Name |
Price |
| 738_KEYp_danioBAC |
Zebrafish BAC Pilot (Keygene) |
£3,545.00
|
The DNA was partially digested with HindIII. The DNA was cloned into the pIndigoBAC-536 vector, in E.coli DH10B.
Library Information:
Species: Danio rerio
Library: Daniokey Pilot
Strain: Tuebingen
Sex: male
Tissue: testis
Host: E. coli DH10B
Vector Type: BAC
Vector: pIndigoBAC-536
Vector cloning site: EcoR1
While pIndigoBAC-536 is a widely used vector for BAC libraries from a huge variety of species, the actual sequence has not been published. According Luo et al. *
“pIndigoBAC536 has the same sequence as pBeloBAC11, except that the internal EcoR1 site was destroyed so that the unique Eco R1 site in the multiple cloning site can be used for cloning, and a random point mutation was selected for in the lac Z gene that provides darker blue colony color on X-gal/IPTG selection. The GenBank accession number for pBeloBAC11 is U51113."
*Luo, M., Wang, Y.-H., Frisch, D., Joobeur, T., Wing, R. A., and Dean, R. A.
(2001) Melon bacterial artificial chromosome (BAC) library construction using
improved methods and identification of clones linked to the locus conferring
resistance to melon Fusarium wilt (Fom-2). Genome 44, 154–162.
Protocols / Clone Handling
Plates: contain LB broth with 7.5% glycerol and chloramphenicol. These are sent on Dry Ice. These should be stored at -70°C.
Clones: are streaked onto LB agar containing chloramphenicol (12.5µg/ml).
To use: re-streak the clone onto the same agar type, in such a way as to obtain single colonies. Incubate at 37°C overnight. Clones can be grown in LB broth + chloramphenicol, and glycerol added after growth to a concentration of 25% for long-term storage at -70°C.
References
Suster, M.L., Abe, G., Schouw, A. and Kawakami, K. (2011). Transposon-mediated BAC transgenesis in zebrafish. Nat Protoc 6, 1998-2021
Howe, K, Clark, M.D. et al (2013) The zebrafish reference genome sequence and its relationship to the human genome Nature 496 (7446), 498-503
Subsequent publication:
Please acknowledge the originators of the library, R.Plasterk and Keygene N.V., and Source BioScience for providing the clones in any publication arising out of using this resource.
For further information and prices please contact us or call +44 (0)115 973 9012