I have prepared a series of diluted template to be used to generate standards but I have not yet add in mastermix as our thermocycler is booked fully so I planned to run the samples later. Can that be kept in 4C fridge or should be kept in -20C for awhile? Anyone who thinks it's fine to keep samples for QPCR before running it? How long should I keep my sample since I haven't added mastermix into the reaction? Thanks
Hi Nurul Kamal, it is always better to keep your cDNA samples frozen at -20°C. Avoid repeated freeze/thawing cycles.
In our lab, we kept diluted cDNA samples at -20°C. On a few occasions, we kept the qPCR plate at 4ºC (template + master mix) for less than 1 hour when the Thermocycler are busy.
Never tried it before, but I'm sure if it's DNA you will not have any trouble keeping it at 4 °C ON.
For qPCR standards I use plasmid DNA, I make serial dilutions final volume 100 ul and then make aliquots of 5 ul (the reaction volume). Therefore, I only take them once out of -20°C, avoiding freeze/thawing cycles.
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