Scientists use qPCR in basic research and clinical diagnostics to measure gene expression, pathology (especially infectious disease and oncology), genetic modification in organisms, and microbiologic ecology. Because of how popular qPCR is, many different protocols, reagents, kits, and products have been made for it. Many parts are made to work best for certain uses, systems, and instruments. All of this variety is good for science, but it can make everyday experiments harder to do.
Most users focus on putting together the PCR reaction solutions and getting the template nucleic acids, but it’s important to consider the PCR plate. Make sure you have the right plate since the wrong choice can affect the quality and consistency of your results.
Moreover, these plates are sealed with highly specialized material sealing. Like AF-100 by Molecular Biology Products is the ideal aluminum foil sealing for PCR plates for long-term storage, light-sensitive assays, and robotics handling.
Things To Know For Choosing The Right PCR Plates
Here are some of the vital things to consider to help you choose the right plate.
1. How many wells are needed, and should they be divided?
This will depend on how big your experiment is (how many reactions you need to do) and what your qPCR machine can do. You may need 96-well plates, 384-well plates, or both as a good place to start. If your qPCR machine can read either format, you might want to have both on hand.
There are also 24-well and 48-well plates, but they aren’t used as often as 96-well and 384-well plates. Some people choose these formats for experiments that don’t need 96 wells. You can use these plates as full 96-well plates or break them up into 32-well, 24-well, or 8-well sections. Plates that are broken up into smaller pieces use less plastic and cost less, and they can be used in more ways.
2. Does every plate work with every instrument?
Not all thermocyclers, qPCR instruments, robotic arms, or samplers can fit all plates. The skirt, deck, well rim, and notch are the four main parts of a PCR plate that determine how well it works with other plates.
The panel that goes around the plate wells is called the “skirt”.Its main job is to keep its shape so that it can be handled and pipetted. Even though non-skirted PCR plates can be used with many thermocyclers and qPCR devices, they can’t be handled by robots.
Types of Skirted Plates
There are two kinds of skirted PCR plates: those with a half skirt and those with a full skirt. Both types of plates add more support to the structure and can be moved by robots, but full-skirted plates are made for instruments with raised blocks and have more mechanical strength. Because robotic arms can bend or warp other plate formats more easily, fully skirted plates are the best choice for automated workflows.
3. What is the footprint of ANSI/SBS?
If you are using high-throughput systems, you should buy plates that fit the ANSI/SBS footprint, which was set by the American National Standards Institute (ANSI) and the Society for Biomolecular Sciences (SBS). The ANSI/SBS footprint was made to use PCR plates with different robotic handling systems.
4. How are clear wells and white wells different?
Users should choose white wells instead of the usual clear ones for qPCR reactions. White wells make assays more sensitive and reduce the amount of variation in qPCR data by stopping fluorescence from getting into the thermal block. They also reduce the amount of fluorescence signal refraction. Because of these advantages, more fluorescence returns to the detector.
Microbio procedures’ results depend on the use of the right products and materials and, of course, the handling methods. The new advancements in PCR plates have revolutionized the procedure of PCR. There are different kinds, like skirted and non-skirted PCR plates. The choice of either of them rests on their usage in the lab.