A target is the specific genetic element (an introduced DNA sequence, a point mutation, or and introduced targeted element (lacZ, Neo, GFP)) which we are amplifying and detecting using a PCR-based method. Each allele that has been modified in a line is a separate target.
The basic unit of heredity that occupies a specific location on a chromosome. Each consists of nucleotides arranged in a linear manner.
A target is the piece of DNA we are amplifying, and the gene is the location.
A transgenic animal has had foreign DNA deliberately inserted (usually at random) into its genome. Transgenes can be detected by both qualitative and quantitative methods depending on customer needs-this need is largely driven by the breeding scheme (see image below), but transgenics under development may also require the use of quantitative PCR.
A knockout is a laboratory mouse in which researchers have inactivated, or “knocked out,” an existing gene by specifically inserting a construct designed to stop its expression. Very commonly this can be a “neomycin insertion” into a critical exon of a gene-but other elements are used as well-hygromycin, lacZ.
A targeted mutation is very similar in method to a knockout. A specific genetic element is introduced to a gene at a specific known location. This element may or may not affect the expression of a gene. Some examples of targeted mutations may be insertion of DNA sequence into a mouse gene to “humanize” the gene (i.e., make the mouse gene more human like). Or the insertion of loxP sites into a gene, that will allow for the conditional knock out of a gene after exposure to Cre Recombinase. We also test for Knock-Ins, SNP’s and INDELS that are produced by genetic modification.
We routinely test mouse, rat, and zebrafish samples, but we can accept certain other species. Please contact us to inquire.
Create an account to be able to submit sample requests online and receive account and request summaries. If you will not be submitting online, simply download and submit a completed genotyping request form with your samples. Email us to let us know they are on the way. For complete details, see the sample submission page.
Yes, samples can be sent at room temperature. In fact, we prefer samples be collected and shipped at room temperature. Samples that are frozen and then are allowed to thaw will experience significant degradation. If you absolutely must freeze the samples, please make sure they remain frozen at all times, including during transit. In the picture below, samples 53 54 and 55 had been previously frozen and allowed to thaw prior to shipping. Samples 52 and 116 demonstrate intact genomic DNA.
Yes, strains that added to your account are only visible to you.
Known controls are only required for validation of new assays. We automatically save control samples at our facility, so you do not need to send them with every order.
DNA is not preferred, but we can accept it if we know the method used to make the DNA. Spin column kits tend to provide much lower yields which can lead to insufficient DNA for genotyping, especially for validations or when multiple targets need to be tested. We do charge for “No Calls” when the DNA has been provided to us.
If using Pro K digest, send 20-50ul, if using a spin column or magnetic beads send 50-100ul. Ship in screw top tubes (preferred), plates with cap mats, or tubes covered securely with paraffin to prevent leakage and cross contamination. Samples can be shipped frozen or at room temperature.
Yes, we can accept frozen embryos. Each embryo needs to be in a separate well or tube. We do not separate multiple tissues together in a single tube or well and we do not resample submitted tissues larger than 2mm.
Yes, however we only accept samples equivalent to a 2mm section of tail. Larger pieces of tissue are likely to fail, and we will not dissect samples into smaller pieces. Organ samples must also be shipped frozen. We do not accept samples which have been placed in a preservative.
No. Larger samples are likely to fail, and a surcharge will be applied.
We offer our design expertise at NO CHARGE, but known controls must be provided.
If the strain is commercially available, send us an email and we will look into it. We will let you know within 24 hours if we can do the testing. We may already have all the necessary primers, but not have the strain built into our system. If we do not have the necessary targets/alleles available, there is a link in your account to a submission page for submitting new targets for assay designs. We offer assay designs free of charge.
You can send them at any time. If the primers/probes have not yet arrived, we will freeze the DNA until all the necessary supplies are present.
We prefer a het and hom for targeted mutations and a hemi and hom for transgenic zygosity. However, if both are not available, send what you have. Without hom controls we cannot confirm our assay will detect homs until one is detected.
This is helpful but not always necessary. Genetic background is important if you are working on a less commonly used inbred or outbred and we need to design primers specific for a wildtype sequence. For example, the sequence of a given gene for C57BL/6J may be very different compared to a wild derived strain such as PWK.
Hydrolysis probes are probes that are hydrolyzed during the amplification process, releasing the fluorophores to allow detection of amplified products. They offer the advantage of an extra layer of specificity for target detection.
SYBR green is one of several intercalating dyes that we use that allows real time detection of product amplification. When combined with melt curve analysis it provides a higher level of specificity.
We guarantee delivery of your results within 2-3 business days of receiving your samples. Validation of new assays will take longer and additional control material may be requested.
We use the following terminology to describe sample results.
- Wild= (Wild-type) for KO (knockouts) and KI (knock-ins), these mice lack the mutation that is being tested for by the assay.
- Hemi= (Hemizygous) for transgenics, these samples have the transgene on one of the pairs of homologous chromosomes but lack the transgene on the other. Used for qPCR results.
- Het= (Heterozygous) for KO and KI, these samples have the mutation on one of the pairs of homologous chromosomes and have wild type sequence on the other.
- Hom= (Homozygous) for both transgenic and KO and KI, these samples have the mutation/transgene on both homologous chromosomes.
- Negative= for qualitative transgenic assays. Indicates that the transgene is not present.
- Positive= for qualitative transgenic assays. Indicates that the transgene is present.
Please view our sample report page to see our reporting layout. Results are sent by email as they are completed. In addition, this file will be in your account, available for download at any time.
Multiplexed assays use multiple sets of primers in one tube to amplify both the wild type and the mutant targets for the method. The biggest impact on the appearance of your results is that the tm for single peaks (EITHER Wild or Homozygous mutant) will be represented as tm1. Only Heterozygous samples will have a tm2.
Separated assays will utilize two wells for each method. One tube will have only wild type primers and the other tube will have only mutant primers. On the report the results for the two tubes will be separated into two heavily lined boxes. As a result the data will look different. Wild type and Heterozygous samples will have a tm1. Homozygous mutant and Heterozygous samples will have a tm2.
Yes, we accept credit card payments. Your invoice will contain a link to pay securely by credit card. Simply click on the link and follow the instructions.