Hi,
I have a real-time PCR assay plate that is ready to run (SYBR green dye, primers, enzyme, template...everything ready).
But just prior the run, our instrument suddenly broke down (with an unresolvable hardware error). Repair is now pending.
Would you mind sharing (1) how long and (2) in dark at what temperature have you kept a 'all-ready' real-time PCR plate before it can be run?
Please shed some light on this. Appreciated!
I would agree with Debabani on this question. I have stored the foil wrapped sealed plates at 4 degrees C for about 24 hour period will out problems. At the time I was running multiplex (VIC and FAM labeled) TaqMGB assays on a ABI 7900HT system.
As previously mentioned, I have also stored overnight and for 24h Q-PCR plate all ready to go, at 4 degrees C wrapped in foil. I was using KAPA SYBR Green or IQ SYBR Green (Biorad) and the Q-PCR machine Biorad CFX96.
You can give it a try when your Q-PCR is fixed. If you are waiting for more that 48h, let us know if the Q-PCR worked that would be helpful to know how long we can store a plate.
Dear Kay,
Just my two cents: I guess it depends a lot on what (high quality) PCR chemistry you are using. I stored a plate for four hours in a dark cooling chamber at 8 degress with only some foil on top of it and the efficiency and overall performance of the run was greatly decreased (2 - 3 cycles higher than usual, less fluorescence in general). If you know how your assay should look like overall you could be fine. If you tried something new you definitely have to repeat it.
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