How to arrange the ideal PCR spot in the lab

First of all, you need a space in the lab of a sufficient size to place the basic equipment needed to prepare a PCR. Depending on the size of the thermal cycler at least 1,20 m should be reserved, so that you have sufficient space to work. Therefore, thermal cyclers with a small footprint and air vents in the back make sense. We recommend having everything near by so that you don’t have to stand up during preparation. If possible, the freezer with the reagents should be below, or next to the bench you chose for your PCR spot. Like this samples are transported a minimal way and constant cooling is guaranteed. If your lab does not have that much space, PCR cooler and ice buckets are mandatory to keep the enzyme inactive and the samples cool until your PCR run starts.

Basic equipment:

• Thermal cycler
• Tip boxes with tips (10 µL, 100 µL, 300 µL and 1.000 µL)
• Pipettes (10 µL, 100 µL, 300 µL and 1.000 µL)
• Cleaning wipes and cleaning agent
• Tube rack
• Sterile tubes to prepare the Mastermix
• Sterile PCR tubes/tubestrips or PCR plates
• Self-adhesive film/foil to cover the plates
• Ice bucket
• Garbage bin

With the basic equipment you will be able to prepare a good PCR experiment when considering some behavior rules to keep the place and your samples clean. It is important to wear a clean lab coat and sterilize your hands thoroughly before pipetting. You should also clean the place before every PCR set-up with the cleaning agent (e.g. DNA Away^® ). Also, the pipettes and tip boxes should be cleaned regularly.

Challenging PCR needs different equipment

If you have many samples and mainly use PCR plates you might also consider some more equipment to facilitate your PCR preparation. We call it the “master equipment” and explain the benefits in every bullet point.

Additional master equipment:

• Multipette ( manual or electronic ): This multi-step liquid dispenser helps you when many tubes, or wells of a plate must be filled with the same liquid, e.g. Mastermix. You can aspirate the full volume at once and then dispense the volume needed in each well with only one movement, or automatic with the electronic version. Dispensing speeds up your preparation and reduces stress on your arm muscles.
• CombiTips: These tips are needed for the Multipette. We recommend the volume sizes 1 mL to fill a full plate with 10 µL Mastermix each well.
• PCR cooler: These cooler changes the cooler when it gets too warm to make you aware of your samples condition. It is ideal to place a full plate or PCR tubes/tubestrips inside while pipetting.
• ThermoStat C with SmartBlock 1.5 mL: The thermal heater/cooler can be cooled down to 4°C to maintain a stable and more reliable temperature in comparison to an ice bucket when cooling the enzymes during PCR preparation.
• HeatSealer and heatsealing foil/film: This method of sealing PCR plates is tighter than self-adhesive sealing film or foil. It is the highest level of evaporation protection for your samples. The foil can be pierced afterwards from most automated liquid handling systems for downstream processes.
• Centrifuge: a centrifuge that is only used for PCR samples is only recommended in laboratories with very high sterility and documentation levels. Most of the time any centrifuge in the lab can be used to spin down the samples after these have been tightly closed.
• PCR workstation: This workstation is used to cover the complete PCR preparation area and only your hands enter it while preparation. The function is comparable to a sterile workbench, but often smaller. DNA denaturation also works via UV light in most cases. Ideally you don’t remove the hands during the whole preparation process. Usually, the thermal cycler is not placed inside the workstation and the closed samples are carried to the device.
  
  

No matter if you decide to start with the basic equipment or if you enhance your PCR spot with some components from the master equipment, always keep the most important point “cleanliness” in mind to prepare comparable, reliable and satisfying PCR results.

Watch now how to set up the ideal PCR spot in your lab: