DryMicro MPXV qPCR Kit

Simple workflows: All-in-one solution with all components included

Easy to store: The master mix can be stored at room temperature for over one and a half years without compromising quality

Improved stability: Freeze-drying increases long-term stability and consistency in product performance

Enhanced flexibility: This kit can be used with purified MPXV DNA or with virus lysed in our proprietary ExtractFree™ buffer (#IV100)

Detection without DNA extraction: One of the remarkable features of this product is that you do not need to extract DNA before performing a PCR. Simply lyse MPXV in ExtractFree™ buffer (#IV100), add the lysate directly to the lyophilized master mix, and you can run real-time PCR the same way you use purified DNA.

One and a half years if the package is not open.

• DryMicro™ PCR Master Mix (Cat# DMD10: Lyophilized microsphere containing thermostable DNA polymerase, MgCl2, dNTPs, optimized PCR buffer, and a proprietary mixture of lyoprotectants.
• Primers and probe solutions: Primers and probe are supplied in separate tubes. The probe is labeled with FAM as the 5’ reporter dye and ZEN / Iowa Black FQ as the 3’ quencher. Both primers and the probe target MPXV F3L gene.
• ExtractFree™ buffer (#IV100): The buffer is used to lyse the virus and release its DNA. The buffer can be added directly to the lyophilized master mix without the need to extract the DNA first.

Store DryMicro™ PCR Master Mix (Cat# DMD101) and ExtractFree™ buffer (#IV100) at room temperature. Store primer tube and probe tube at -20 ℃.

• Add 17 µL of nuclease-free water containing viral DNA or ExtractFree™ buffer (#IV100) with the viruses to the DryMicro™ PCR Master Mix (Cat# DMD101) tube containing lyophilized microsphere, vortex for 5 seconds and spin down the solution using a micro centrifuge. The lyophilized powder shall dissolve immediately.
• Add the forward and reverse primers and/or probe(s) to the rehydrated master mix, bring the total volume of the mix to 20 µL with nuclease-free water. Vortex for 3 seconds and centrifuge briefly to make sure all solutions are collected at the bottom of the tube.
• Load the tube onto a real-time PCR detection system. If you use a 96- or 384-well plate, you can transfer the mix into the designated wells.
• Perform PCR according to the following program:

                 Step 1: 95ºC, 2 minutes; 1 cycle

                 Step 2: 95 ºC, 5 seconds, then 60 ºC, 10 seconds; 45 cycles.

MPXV101

Monkeypox virus (MPXV) is a member of the orthopoxvirus genus. The MPXV genome consists of a double-stranded DNA, approximately 197 kb in size and coding about 190 proteins. Currently, real-time PCR (qPCR) is a standard laboratory and diagnostic technique for the detection of MPXV. However, most commercial kits come in solutions that require a cold chain for transport and freezers for storage. To facilitate transport and storage, we optimize the enzyme and buffer components of the kit and lyophilize them in the form of microspheres. With the addition of solution, the lyophilized microspheres dissolve instantly without sacrificing their original qualities. Not only does the rehydrated mix match the performance of established brands, but it can also be used for the detection of pathogens without the DNA extraction step, thus greatly reducing time and costs.

• If you choose to run your qPCR without using our probe, you can use your own fluorescent dye such as SYBR Green or purchase our fluorescent dye (Cat# FL110) which can be detected at FAM fluorescent channel.
• If you choose to use purified DNA in nuclease -free water instead of using our IntactViral™ buffer (#IV100), we recommend that you use Qiagen QIAamp DNA Mini Kit to extract the DNA of MPXV and elute the DNA into nuclease-free water rather than the elution buffer provided.