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Ħ§Time
is money!Ħ¨ 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.
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