“Time is money!” This is an adage everyone agrees.
According to the situation we observed in training courses held in different countries, most technicians had already learned to perform multi tasks at the same time. Due to the familiarity with IQ2000TM kit, everyone understands that PCR premix needed to be thawed in advance before using and this step takes approximately 15 minutes. By saving these 15 minutes, technicians would withdraw a whole kit from freezer and allow all the components in it to defrost in room temperature while performing the extraction at the same time.
However, technicians might have underestimated the time needed for a complete extraction procedure. When it is time to use the solution, kit would have been rested in room temperature for at least one hour. Leaving components in room temperature for too long would lead to the following drawbacks,
1. Reducing enzyme activity: The activity of IQzyme DNA polymerase will decrease though it is a thermal stable enzyme, even RT enzyme is more sensitive to the temperature.
2. Components in the PCR premix decay gradually.
In order to maintain the IQ2000TM Viral Detection and Prevention System at its best condition, following procedures are recommended in order,
1. Take out the same quantity of the PCR premixes and the plasmid DNA standard from freezer equivalent to those of your samples once the DNA extraction procedure has been finished.
2. Thaw completely by the heat of hands or room temperature.
3. Vortex briefly and spin down.
4. Take out IQzyme DNA polymerase and RT enzyme from freezer and spin down.
5. Prepare both first step and nested PCR reaction reagent mixtures.
6. Put the enzyme and premixes back into the freezer.
7. Perform the first step of PCR (or RT-PCR)
8. Store the nested PCR reaction reagent mixture in 40C refrigerator or room temperature during the performing of first step PCR. Make sure it is not stored in 00C or below.
With extra care and attention, kits can be maintained at their best quality and results can be obtained accurately within their shelf life.