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Mouse MICER Resources

Mouse MICER Resources

Source BioScience are happy to announce the distribution a second, fully indexed mouse resource, constructed in Alan Bradley′s lab, The Wellcome Trust Sanger Institute, Hinxton, UK.

Library specifications

The library comprises of more than 93 thousand, constructed, insertional targeting vectors from two different libraries MHPP & MHPN. Approximately 6 thousand of these vectors can be used directly to inactivate genes, with an average targeting efficiency of 28%.

By using the combination of MHPP & MHPN vectors, 3′ Hprt and 5′ Hprt respectively, the disruption of both alleles of a gene may be analysed. Furthermore, larger genomic changes may be engineered, such as deletions, duplications translocations or inversions.

Each clone is a ready made targeting vector, incorporating a drug resistance gene for drug targeting, the 5′ or 3′ portion of the Hprt mini-gene, a loxP site for chromosome engineering. Finally, a mouse coat-colour marker, for stock maintenance is a present on each clone.

MICER clones are displayed on the Ensembl genome browser within a DAS (Distributed Annotation Server). End sequenced vectors are mapped to the C57BL/6J mouse genome assembly. 3′Hprt [MHPP] clones are displayed as blue or green, whilst 5′Hprt [MHPN] clones are displayed as black or red.

Please note, the backbone vectors p5′ Hprt [MHPN] and p3′ Hprt [MHPP] are also available at Source BioScience.

If you would like further information regarding this resource, please go to the following URL: www.sanger.ac.uk/PostGenomics/mousegenomics/

Source BioScience offers the complete clone set of 821 384-well microtitre plates, or individual clones, for research purposes only.

Protocols / Clone Handling

Plates contain LB broth with 7.5% glycerol and ampicillin. The plates are sent on Dry Ice, and should be stored at -70°C.

Individual clones pools have been streaked onto LB agar containing ampicillin (50 μg/ml). Please store them at 4°C (not in a freezer). As soon as possible, transfer the culture to LB broth containing ampicillin + 7.5 % glycerol and incubate overnight at 37°C (do not try to transfer single colonies, instead scoop out the whole bacterial culture using a disposable inoculation loop). Incubate overnight at 37°C for subsequent freezing at -70°C.

Bacterial strain DH10B genotype: F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara leu)7697 galU galK rpsL endA1 nupG.

References

Mutagenic insertion and chromosome engineering resource (MICER). Adams DJ, Biggs PJ, Cox T, Davies R, van der Weyden L, Jonkers J, Smith J, Plumb B, Taylor R, Nishijima I, Yu Y, Rogers J, Bradley A. Nat Genet. 2004 Aug;36(8):867-71. Epub 2004 Jul 4. The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambs, CB10 1SA, UK.


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