Molecular Diagnostics Services, Inc.
A Contract Research Organization

Transfection


MDS offers a range of services that enable sensitive and accurate determination of gene expression levels. The outline of transfection and purification process that we follow is summarized below.

Vector construction: The desired cDNA is cloned into a mammalian expression vector that carries the gene for neomycin resistance. The construct is fully sequence verified and a large-scale preparation of plasmid construct DNA is made.

Transfection: The construct is incubated with a transfection reagent and transfected into a host cell, typically COS-1, CHO KI or HEK 293 cells. Cotransfection of the vector services are also available. The services include: Insert confirmation: Verification of the transfer of the desired insert into the vector is performed by PCR analysis. Verification of the expression with the client: Both cells and supernatant from plaque-purified recombinants are sent to the client for analysis. We recommend Western or ELISA assays to determine if the protein of interest is present.

Selection and Enrichment: Cells that carry the construct are selected using neomycin resistance carried by the expression vector. Clones expressing the target protein are selected using Quantitative PCR and/or Western blotting. Clonal selection is performed by dilution in 96-well plates and subsequent propagation of clones of interest.

Deliverables: A final cell line with the desired characteristics is selected. Mycoplasma testing is carried out on all clones prior to shipping worldwide.

Additional Services: MDS's Technical Services Division offers many additional services to save you valuable time and resources. Additional services include:
| Home | About Us | Services | Newsletter | Careers | Forms | Contact | Search | Site Map |
©2007 Molecular Diagnostic Services, Inc.