Speed Congenics Service Details

We suggest the following breeding strategy for optimal results. Submit DNA to Van Andel Research Institute after breeding each generation for SNP genotyping, analysis, and reporting to determine "best" male selection for breeding the next generation.

1. Obtain a female mouse that contains the genetic region of interest, such as a transgene or gene-targeted allele. This is the donor strain.
2. Generally, we recommend fixing the sex chromosomes, except for sex-linked mouse models.  
   
   Cross the female donor to an inbred male mouse of the recipient strain.  
   - The inbred recipient strains available for this service are C57BL/6, 129, FVB, BALB/c, DBA, NOD, CBA, AKR, C3H, or SJL.
   - The resulting offspring will be the F1 generation.
   - The Y chromosome in the male F1 offspring will be derived from the recipient strain.
   
   Cross male F1 mice containing the genetic region of interest to females from the recipient strain.  
   - The male N2 offspring will have X and Y chromosomes derived from the recipient strain.  
   - In all subsequent generations, the X and Y chromosomes will be fixed.
   
3. Genotype the N2 male mice for the gene of interest.
   
   Submit the DNA of positive genotyped N2 male mice for VARI speed congenics service.    
   
   The report will indicate the percentage of recipient background in each mouse sample.  The “best” male to mate will be the one with the highest recipient genomic background. 
   
4. Backcross the “best” male of N2 to the female recipient strain to produce N3 animals.
5. Submit the DNA of positive genotyped N3 male mice for VARI speed congenics service to select the “best” male, and then mate with female recipient strain to produce the N4 generation.
6. Repeat step 5 with positive genotyped N4 male mice, submitting their DNA for the “best” male selection to produce the N5 generation.
7. At the N5 generation, more than 99% of the genome should be derived from the recipient strain background.

DNA Submission

1. Please complete the Speed Congenics Service Project Application Form and e-mail this to Sok-Kean.Khoo@vai.org or fax to 616-234-5537.
2. Van Andel Research Institute (VARI) requires a signed service agreement prior to initiating any project. The service agreement will be sent to you prior to starting the project. Once the agreement has been completed, signed and sent back to VARI, we will begin the project.
3. Mouse tail DNA is required.  The DNA concentration should be at least 75 ng/µl in a total volume of at least 20 µl.  
   
   Sample number should be in batches of 16 for the 16-sample array, and at least 46 samples should be sent for the 96-sample array to be cost effective.  
   
   For 16-sample array, if you have two congenics projects at one time, you may combine the two groups of samples (approximately 8 each) into one batch to fit into one array.
   
4. Please ship your samples on dry ice via overnight courier to:

Sok Kean Khoo, Ph.D.
Laboratory of Germline Modification and Cytogenetics
Van Andel Research Institute
333 Bostwick Ave., N.E.
Grand Rapids, MI 49503

Phone: 616-234-5536
Fax: 616-234-5537
E-mail

Mouse SNP Genotyping, Analysis, and Report

1. DNA is genotyped on the Illumina’s Mouse Medium Density Linkage Panel consisting 1,449 SNP loci, using the Illumina GoldenGate Assay and the Illumina BeadStation.
2. SNPs in this panel provide a uniform genome distribution at a density of approximately three SNPs per 5-Mb intervals across the whole genome.  Additional Information
3. Both homozygous and heterozygous alleles of the DNA are scored according to the recipient genomic background. The ranking of the “best” male is according to the highest percentage of the recipient genome.
4. The turnaround time is approximately 2 to 3 weeks following receipt of DNA samples.
5. A report that includes the ranking of the “best” male will be sent via e-mail upon completion of the analysis.