Genotyping

Genotyping

On the one hand, genotyping is used to clarify whether a laboratory animal is transgenic or not. On the other hand, it can be determined whether the animal is homozygous or heterozygous and how many copies of a gene are present. The method of choice here is PCR and RT-PCR.

We perform single PCRs as well as double or multiple allele PCRs. We offer restriction digestion and gel electrophoresis as well as RT-PCR and sequencing for SNPs and indels.

The already established PCR assay is provided by the customer or standard PCR assays (Jackson Lab) are used for porting, validation and optimization. In addition, we offer the development of new PCR assays.

As little as 2-3 mm of a tail or ear biopsy is sufficient for such testing.

The pre- and post-PCR areas are clearly separated in the laboratory to avoid contamination. Additional control tests that run with each assay ensure this. Lysis and DNA extraction of the biopsies is performed in 1.5 ml reaction tubes, amplification in PCR (Applied Biosystems, Biometra, peqlab) and RT-PCR cyclers (Applied Biosystems).

The number of our PCR cyclers allows us to process more variations of PCR programs simultaneously than would be possible with a gradient cycler. By using 12 PCR cyclers, not only temperatures but also times can be varied, thus saving time in the optimization of PCR assays.

Gel-based detection of PCR products is performed by high-throughput capillary gel electrophoresis using the QIAxcel system (Qiagen). Deletions/insertions and SNPs are sequenced in special cases by the 3730 DNA Analyzer (Applied Biosystems).

We provide these services in cooperation with StarSEQ GmbH, Mainz.