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Quality Control
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The genetic integrity and health quality of RIKEN BRC mice are guaranteed
by extensive quality control programs. High-quality mice are distributed
to users in the scientific community.
1. Microbiological Monitoring
1) Sentinel mouse program
The RIKEN BRC has adopted the sentinel mouse program, a specialized approach
with the microisolator cage system. A sentinel mouse cage containing five
mice is laid on a microisolator cage rack and small pieces of bedding material
from all cages of the rack are collected and placed in the sentinel mouse
cage. Two mice from a sentinel cage are subjected to examination quarterly;
one is for in-house examination and the other is sent to the ICLAS monitoring
center of the Central Institute for Laboratory Animals, Kawasaki, Japan.
The health report you receive is a copy of the then-most-recent Health
Certificate guaranteed by a veterinarian at the ICLAS Monitoring Center.
We classify the microorganisms into four classes according to the risk presented
to the mouse in Japan. All sentinel mice are tested for class A and B. Sentinel
mice kept in the room for immuno-deficient mice are screened for class C as well.
Class D contains certain pathogens believed not to exist in the Japanese mouse colony;
therefore, mice are examined only at the request and expense of the recipient.
The microbiological environment of our facility is also monitored periodically for
bacteria and fungi.
BRC facility Health Certificate 2010.4-2011.11
Class A:
Clostridium piliforme (Tyzzer's organism), Ectromelia virus, Hantaan virus, Lymphocytic choriomeningitis virus (LCMV), Mouse hepatitis virus (MHV), Mycoplasma pulmonis, Salmonella typhimurium, Sendai virus (HVJ)
Class B:
CAR bacillus, Citrobacter rodentium, Corynebacterium kutscheri, Dermatophytes, Ectoparasites, Helicobacter bilis, Helicobacter hepaticus, Intestinal protozoa, Pasteurella pneumotropica, Pinworms, Salmonella spp.
Class C:
Stapylococcus aureus, Pneumocystis carinii f.s. muris, Pseudomonas aeruginosa
Class D:
Lactate dehydrogenase elevating virus (LDHEV), Mouse adenovirus (MAV), Mouse cytomegalovirus (MCMV), Mouse minute virus (MMV), Mouse norovirus (MNV), Mouse parvovirus (MPV), Mouse polyoma virus (Poly), Mouse rotavirus (EDIM), Pneumonia virus of mice (PVM), Reovirus type 3 (Reo 3), Theiler's mouse encephalomielitis virus (TMEV)
2. Genetic Monitoring
The Genetic Quality Control Program is to confirm the genetic integrity
of deposited mouse strains in the process of rederivation treatment, colony
maintenance breeding, and preservation of frozen stocks through the following
monitoring protocols.
1) Molecular Genotyping
Molecular genotyping is performed in order to identify and confirm carriers
of induced mutations (targeted mutations and transgenes) and spontaneous
mutations. Genomic DNA from the tail-tip of an individual mouse is extracted
and purified by an automatic DNA extraction machine. Genotyping protocols,
including primer sequences, enzymes and thermal cycling programs for each
strain, are determined from information provided by the Depositor. In some
cases, protocols are modified for more efficient genotyping.
2) Genetic Background Monitoring
For the surveillance of the genetic integrity of mice strains in the RIKEN
BRC, we use the following assays for genetic background monitoring at critical
points in the process of colony maintenance and preservation.
i) Electrophoretic testing for biochemical markers
Fifteen biochemical markers out of 19 standard genetic markers in the ICLAS
monitoring center are examined to profile the genetic background of the
strains. Most of these assays are tests of isoenzymes through the use of
cellulose acetate membrane or SDS polyacrylamide gel electrophoresis. Samples
for the assays are kidney homogenate, serum, erythrocyte lysate and urine.
The genetic background of the strain is identified by comparing the result
with published standards for inbred strains.
ii) Microsatellite marker testing
We have developed a panel of Simple Sequence Length Polymorphism (microsatellite)
markers that can be used to distinguish between strains. From the survey
of microsatellite markers, we have selected 96 highly polymorphic markers
that represent the relatively large difference in PCR product sizes easily
distinguishable by agarose gel electrophoresis. The RIKEN BRC is in the
process of making the transition from biochemical markers to molecular
markers for genetic surveillance of strains because the latter provide
more information concerning genetic background.
iii) Karyotyping and chromosomal mapping by FISH (Fluorescence in situ Hybridization).
Strains with chromosomal anomalies are examined for their karyotype using
culture cells from the spleen. For transgenic strains chromosomal mapping
of the transgenes are performed by FISH.
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