They should be OK if you did not defrost and freeze them several times when they were in -80! You can simply move them back into -20 if they are frozen in -80.

They should be OK if you did not defrost and freeze them several times when they were in -80! You can simply move them back into -20 if they are frozen in -80.

Make aliquots that are equivalent to X number of reactions to minimize the thaw/freeze. I've done that with enzymes several times and had no issue. Recommend making whatever your typical # of reactions is plus 10 to 15% for some overage.

The problem of repeated thaw-freeze cycles would impact the quality of your reagents much more than the storage temperature.

Real time PCR Reagents not allowed to keep in -80 it will effect on ur PCR results so u can remove reagents from -80 and keep it in zero degree centigrade normal fridge also .

This causes no problem at all.

It could be dependent on the brand of your reagents. Some reagents seem to be stable for quite long periods also when stored in the fridge. Other reagents showed a decrease in efficiency after 3 days of storage at temperatures above -20. I have not found any information about the storing at -80 degree.

Here are the information that I found:

An RT-PCR kit developed by the Medical Entomology Unit, Institute for Medical Research to detect dengue infection in mosquitoes, was tested for its shelf life at 3 storage temperatures: room temperature, refrigerator and freezer. Test kits were tested once every 3 days for kits stored at room temperature, and once every week for those stored at refrigerator and freezer temperatures. The results showed that the test kit could only be stored above its recommended storage temperature of -20 degrees C for not more than 3 days (Cher Pheng Ooi et al. Trop Biomed. 2005 Jun).

KiCqStart SYBR Green qPCR ReadyMix is stable for 1 year when stored in a constant temperature freezer at -20°C, protected from light. For convenience, it may be stored unfrozen at +2 to +8°C for up to 6 months (sigma Aldrich).

Hi Ahmed, it should not be a problem if you avoid repeated freeze and thaw of your reagent.