How is mice genotyping done?
There are several ways to obtain DNA for mouse genotyping. You can collect DNA from ear tissue (0.5 to 2 mm) using an ear punch. This yields a larger quantity of quality DNA. Ear tissue is easy to collect when mice are ear punched for identification.
Which technique can be used for identification of transgenic mice?
The founders are typically identified by either a Southern blot or genomic PCR assay, often using the proteinase-K-digested tail DNA and eventually be used to establish independent lines that will vary with regard to the transgene integration site as well as in its copy number.
What is a primer in genotyping?
Internal control primers typically amplify a genomic DNA region that is unrelated to your gene of interest. Successful amplification of the internal control indicates your DNA is suitable for PCR.
Why do we do genotyping?
Genotyping determines differences in genetic complement by comparing a DNA sequence to that of another sample or a reference sequence. It identifies small variations in genetic sequence within populations, such as single-nucleotide polymorphisms (SNPs).
Why do we genotype mice?
The genotype of your mouse determines the phenotype. Therefore, a homozygous mouse will have different characteristics than a heterozygous mouse. For this reason, it is important to routinely genotype your mice to maintain consistency with your mouse colonies.
What is transgenic mouse model?
Transgenic mice are mouse models that have had their genomes altered for the purpose of studying gene functions. At Charles River, we help hundreds of global customers by delivering study-ready, transgenic mice to meet their research needs.
Why mice are used as transgenic animals?
Such work laid the basis for the creation of transgenic mice genetically modified to inherit particular forms of cancer. These mice were generated as a laboratory tool to better understand the onset and progression of cancer. The advantage of such mice is that they provide a model which closely mimics the human body.
What is mouse genotyping?
Genotyping is the process of detecting the presence, or absence, of specific DNA sequences in a particular organism’s genome. Since genes can influence a mouse’s phenotype, being able to probe an individual mouse’s genetic make-up, or “genotype,” is critical for attributing a phenotype to a specific gene.
What is PCR genotyping?
Genotyping PCR is used to determine the genotype of an organism (e.g., WT vs. mutant, or WT vs. transgenic). PCR primers are designed to specifically amplify either a portion of the transgene (in a transgenic animal) or the mutation (in a mutant animal). They are then used in a reaction containing DNA from the animal.
What is genotyping PCR?
What kind of transgene is used in mouse genotyping?
The most common “transgene” introduced into mice is one that drives the expression of green fluorescent protein, or GFP, from jellyfish. By using a tissue-specific promoter (a regulatory sequence that “promotes” the activity of a gene) to drive GFP production, cells from a specific tissue type can be easily identified by green fluorescence.
Why do you need A Jax genotyping protocol?
You have determined that you need a JAX qPCR genotyping protocol so that you can distinguish between hemizygotes and homozygotes for a transgene. Let’s take a look at a qPCR protocol for the Tcra transgene in the OT-I transgenic mice as an example of what kind of information you can find on a JAX qPCR genotyping protocol.
How is a band detected in a transgenic mouse?
A band will be detected for the endogenous mouse gene in all samples, and transgenic mice will be indicated by the presence of an additional band. The detection of a band in all samples serves to control for DNA quality and correct amplification.
How can PCR be used for mouse genotyping?
Here we describe polymerase chain reaction (PCR) genotyping protocols for fast, sensitive, easy, and cost-effective characterization of mouse genotype. We discuss optimization of parameters to improve the reliability of each assay and propose recommendations for enhancing reproducibility and reducing the occurrence of inconclusive genotyping.