How to Run Multiple Genotyping Tests In Parallel

Many tests and one bundle to rule them all!

The strength of miniPCR Dx gelPCR bundles is that a single bundle opens the door to countless genotyping possibilities. Whether you’re a ball python breeder screening new hatchlings for a range of recessive traits, or a passionate fowl breeder selecting for sex and desirable traits within the same bird, miniPCR Dx empowers you to achieve more with less, all with a single, cost-effective equipment bundle.

What do you need

To run miniPCR Dx gelPCR tests you will need ONE gelPCR bundle and the kits. We have two options, both work great and it just depends on your needs:

• gelPCR™ Starter Bundle: Processes 8 samples (7 tests + 1 negative control).
• gelPCR™ Premium Bundle: Processes 16 samples (15 tests + 1 negative control).

Good news: equipment is a one-time purchase!

How it Works: The Beauty of Simplicity

The convenience of a gelPCR bundle lies in its ability to handle multiple genotyping assays using the same core equipment. While each specific test is uniquely designed and tailored to your target of interest, they all follow a consistent workflow, specifically:

1. DNA Extraction: The crucial first step, isolating the genetic material from your sample (e.g., a scale from a snake shed or a drop of blood from a chick).
2. PCR Amplification: Using specific reagents and precisely controlled temperature cycles within your miniPCR, you create millions of copies of the exact DNA region you want to analyze. Each PCR is designed to target a specific gene.
3. Gel Electrophoresis and Visualization: The amplified DNA fragments are then separated by size on an agarose gel and you can see distinct bands representing the presence or absence of your target genes.

This shared workflow is the key to optimization, allowing you to cleverly screen for multiple genes from that single initial DNA extraction.

Maximum High-Throughput from Your miniPCR Bundles: Working Smarter, Not Harder 

Example One: Setting up a new flock in time for the next show fair

Let’s dive into a practical example. Mary is a chicken breeder and would like to set up a new flock of blue egg layers with lavender plumage, not carrying recessive white, to sell at the next show fair of the year. To speed up the process and make sure she can get the project done and meet the deadline, Mary needs to screen candidates chicks for four traits (sex, blue egg, lavender, and recessive white). How can you efficiently get all this information? Here are three simple yet powerful steps to make your life as a breeder easier:

1. Extract DNA Once: Perform a single DNA extraction from each candidate chick. This extracted DNA now holds the entire genetic information you need for all your tests.
2. Prepare Different PCRs: For each trait you want to test, you’ll set up a separate PCR reaction. Each PCR tube will contain the same DNA extract from a certain chick, but will differ in the specific reagents designed to amplify the target gene for that trait, as well as slightly different PCR settings. The beauty here is that once a PCR is complete, it’s remarkably stable at room temperature! You can confidently let the finished PCR reactions sit on your bench while you prepare the other PCRs and run all the tests sequentially.
3. Run All PCRs on the Same Gel: Once all your PCRs are complete, you can load the products from each reaction into separate wells on the same agarose gel. This allows you to visualize the results for all three traits side-by-side, making interpretation straightforward and efficient. Remember to carefully track which well corresponds to which PCR reaction.

Pro Tips:

• For Power Users: Investing in additional miniPCR units unlocks the potential to run all your different PCRs in parallel, significantly reducing your overall processing time!
• For High-Throughput Heroes: Meet GELATO™, miniPCR’s electrophoresis and visualization system designed for speed and capacity! With GELATO, you can run an impressive up to 50 PCR reactions simultaneously! This means you could potentially screen a dozen chicks for four different traits (e.g., sexing, blue, lavender, and recessive white) in a single gel run! GELATO is included in the gelPCR™ Premium Bundle.

Example Two: Selling a certified ball python by the end of the day

John has posted some pictures of one of his Piebald snakes on his website and there are some very interested customers. A potential buyer is willing to pay $300 dollars more if he knows whether the snake is Het for Lavender Albino or Ghost, but the offer will expire soon. Thankfully John has his own miniPCR lab and he kept the shed of this snake. He goes to the lab and quickly:

1. Extracts DNA Once: Performs a single DNA extraction from the shed, in 10 minutes the DNA is ready.
2. Prepares Different PCRs: Sets up two PCR reactions sequentially, one for Lavender Albino and one for Ghost. In about 3 hours both PCRs are ready.
3. Runs All PCRs on the Same Gel: Once all his PCRs are complete, he visualizes the results for both traits side-by-side, making interpretation straightforward and efficient.

The Sweet Advantages of Optimizing Your Runs: Time and Resource Savings

By adopting this optimized approach to your miniPCR workflow, you’ll reap significant benefits:

• Saving at the DNA Extraction Step: The most labor-intensive and resource-consuming step is often DNA extraction. By performing it only once per sample, regardless of how many genes you want to test, you drastically reduce your hands-on time, the amount of extraction kits you use, and the associated costs.
• Saving at the Gel Electrophoresis Step: Running all your PCR products for a single sample on the same gel minimizes the number of gels you need to prepare and run, saving you time, agarose, buffer, and staining reagents. This also simplifies the analysis and documentation process.

Embrace the power of multiplexing with your gelPCR bundle and unlock a new level of efficiency in your breeding endeavors!

Related Resources (at dx.minipcr.com)

• gelPCR™ Starter Bundle: Processes 8 samples (7 tests + 1 negative control)
• gelPCR™ Premium Bundle: Processes 16 samples (15 tests + 1 negative control)
• miniPCR® mini8X thermal cycler (4-unit bundle)
• miniPCR® mini16X thermal cycler (4-unit bundle)
• Avian tests
• Ball Python tests