Services

• Mouse and Rat Microsatellite and SNP Genotyping Supporting Marker-Assisted Backcrossing (Speed Congenics)
  
• Genetic Background Characterization of Genetically Engineered Mouse Lines
  
• Genetic Quality Control of Inbred Strains 
  
• Mouse and Rat Cell Line Characterization and Mycoplasma Testing
• PCR Testing for Rodent Infectious Agents
  
• H2 (MHC) Haplotyping and PCR Sex Determination
• Passenger Mutation Analysis (e.g., Crb1<rd8>; Nnt deletion, etc.)
• Rodent Genetic Consultation

Mouse and Rat Microsatellite and SNP Genotyping Supporting Marker-Assisted Backcrossing (Speed Congenics)
Transgenic and targeted mutants frequently need to be transferred to a more suitable inbred background for phenotypic analysis or crossed with cre-expressing lines crossed with floxed alleles. Traditional congenic strain development requires a series of backcrosses between the donor strain carrying the mutation (often on a mixed genetic background) and a recipient strain with a defined genetic background (typically an inbred strain) over many successive backcross and taking three years. Marker-assisted (speed congenic) protocols reduce this time from 10 backcross generations to five.

Mouse and Rat Cell Line Characterization and Mycoplasma Testing
Cross-contamination or misidentification of cell lines is a serious problem. It is estimated that 15-30% of human cell lines are misidentified or contaminated, with the consequent invalidation of published data and wasted time and money. Although information on contaminated rodent cell lines is scattered, it is expected to be widespread. LAGS offers SSLP (also known as STR) and SNP analysis for mouse and rat cell line characterization. Upon submission of DNA, we will generate an allelic profile using a standard panel of SSLPs or SNPs. In order to rule out contamination with human or rat cells, we will include human- and rat-specific SSLP primers. It is important to understand that the SSLP or SNP profile will only determine the inbred strain origin for the cell line (and rule out cross-contamination with other cell lines from a different strain or species), but will not identify a particular cell line. For example, examining DNA from a pure B16 melanoma cell line will show a C57BL/6 profile because this is the origin, but the profile will be similar for a LLC cell line, also established from C57BL/6. For ES cell lines characterization, we recommend our tailor-made Background Characterization service.

Contamination of cell cultures with Mycoplasma spp. is recognized as a very serious problem that can significantly impact data interpretation. LAGS uses the MycoAlert™ test from Lonza, a luminescence-based, bichemical test providing rapid screening for Mycoplasma contamination. 

PCR Testing for Rodent Infectious Agents
We offer infectious diseases testing for the following agents: Syphacia obvelata and Aspiculuris tetraptera (pinworms), Myocoptes musculinus, Myobia musculi, and Radfordia affinis (fur mites) Helicobacter spp., Campylobacter spp., mouse parvovirus (MPV)*, rat parvovirus (RPV)*, mouse norovirus (MNV) and mouse hepatitis virus (MHV) and pinworms. Tests for other infectious agents can also be developed and implemented according to the specific needs of your animal colony. 

Passenger Mutation Analysis
Passenger mutations are mutations that are hidden in the genomes of substrains and potentially affect the outcome of an experiment. These mutations typically arise due to genetic drift among separate populations derived from the same original inbred strain, resulting in new and unique phenotypic characteristics. For example, mice of the C57BL/6N substrain (NIH) and not the C57BL/6J (JAX) are homozygous for a single nucleotide deletion of Crb1 (rd8 mutation), which will render these mice inadequate for behavioral studies requiring vision. Other examples include mutations in: Nnt, Skint1, Casp4, Gpr84, Patch1 (susceptibility to SCC), Pde6b (rd1), Bfsp2 (dundee), Gnat1 (IRD1/IRD2), Gnat2 (cpfl3), mt-Atp8<m1> (increased anxiety-like behavior), and Hc (complement C5). In the case of mixed or unknown backgrounds (particularly GEM lines), these mutations can produce unusual or unexpected results. LAGS offers specific genotypes for the mutations listed above.