OmniPCR (OnePCR Ultra) Supermix w Fluorescent Dye

OmniPCR (OnePCR Ultra) is a ready-to-use combined solution containing the Thermostable DNA polymerase, PCR buffer, dNTPs, gel loading dyes, enhancer, and fluorescence dye. Directly add primers, template, and water to execute the polymerase chain reaction (PCR). The Taq DNA polymerase exhibits 5′-exonuclease activity and after carrying DNA electrophoresis, the OmniPCR (OnePCR Ultra) fluorescence dye is directly detected on BluPAD LED transilluminator or other compatible transilluminator. The supermix is provided at 2x concentration and used at 1X concentration for the addition of primer and template solutions. Reagents are provided with the sufficient amplification reactions of 50 μl each, 100 reactions per kit.

• No post-staining processing of DNA required.
• Direct loading onto the agarose gel for electrophoresis.
• Sensitivity: High degree of sensitivity equivalent to ethidium bromide.
• Speed: Benefit for quick set up of PCR reaction such as colony PCR when screening for positive clones.
• Compatibility: Use blue light or UV to detect the signal.

As a starting point, please place the pre-chilled components on ice and follow the below steps:

Standard PCR with OmniPCR (OnePCR Ultra):

• For each 50 μl reaction, assemble the following components in a 0.2 ml PCR tube on ice just prior to use:

OnePCR Ultra	25
Forward primer, 5-10 μM	Variable
Reverse primer, 5-10 μM	Variable
DNA template	Variable
Add ddH2O to	50

• Mix gently. If necessary, centrifuge briefly. Cap tubes and place in the thermal cycler.
• Process in the thermal cycler for 25-35 cycles as follows:

Initial Denaturation	2-5 minutes at 94°C
Denaturation	20-40 seconds at 94°C
Annealing	1 min at the proper annealing temperature
Extension	2 min at 72°C
Final extension	5 min at 72°C

• After the PCR reaction, perform agarose electrophoresis to detect PCR product. No additional dye is required for the PCR samples. Gels can be post-stained with Ethidium Bromide if desired.
• Use the blue-light or UV transilluminator to photograph the gel.

Important notes: OnePCR Ultra is light sensitive and should be stored and protected from light.

Instrument Compatibility

This Super Mix is compatible with the majority of commercially available PCR systems.

• Submerge the PCR product containing the fluorescence dye into the 100 mM NaCl and add 2.5 volumes of absolute or 95% ethanol.
• Incubate on ice for 20 minutes.
• Centrifuge the mixture at 4°C for at least 10 minutes.
• Remove the suspension of ethanol and wash the pellet with 1ml of 70% ethanol.
• Dry the residual ethanol and resuspend the double-stranded DNA in the TE.

• A compatible PCR instrument
• Vortex or equivalent
• Microcentrifuge
• Plates and seals for your instruments