Genotyping

 

Genotyoing is firstly used to clarify, whether a laboratory animal is transgenic or not. Secondly, it can be determined if it is a homo- or heterozygote animal and how many gene copies exist. Here the method of choice is PCR and RT-PCR.  

We conduct single PCRs as well as duplex or multiplex PCRs. Moreover, restriction enzyme digestion, gel electrophoresis, RT-PCR and sequencing for SNPs and Indels are part of our portfolio. An already established PCR assay is therefore provided by the customer or standard PCR assays (Jackson Lab) for transferring, validation and optimization are used. In addition, we offer the development of new PCR assays.

Already 2 to 3 mm of a tail or ear biopsy is sufficient for such a test.

In our laboratories the pre- and post-PCR sections are separated strictly to avoid contamination. This is ensured by additional controls which run along with any assay. Lysis and DNA extraction of biopsies take place in 1,5 ml reaction tubes, amplification in PCR (Applied Biosystems, Biometra, peglab) and RT-PCR cyclers (Applied Biosystems).

Due to our number of PCR cyclers we are able to handle more variations of PCR programs simultaneously as usually possible with a gradient cycler. By the use of 12 PCR cyclers not only temperature but also times can be varied. This gains time in the optimization process of PCR assays.

For the gel based detection of the PCR products a high-throughput-capillary gel electrophoresis, QIAxcel-Systems (Quiagen), is used. In special cases deletions/insertions and SPNs are sequenced with a 3730 DNA-analyzer (Applied Biosystems).

These services are provided in cooperation with StarSEO GmbH, Mainz.