I received fluorescent labelled primers and I am not sure how to re-suspend them. The information sheet doesn't give instructions that are usually given with primers, so I decide to wait until the holiday is over so I can ask a senior lab member to help. But not I am not sure if it is necessary to re-suspend immediately or can I keep them in lyophilized state at -20? I covered with aluminium foil to avoid degradation of fluorescent dye.
Pfano, you can store them in a freezer (-20ºC or better still -80ºC) protected from light for a very long time - decades!
Hi Pfano,
I had lyophilized fluorescently labeled primers for over a month at -20 with no trouble. Just keep them protected from light at -20C, they should be fine. For a shorter period, you can even leave them at 4C.
Good luck!
Pfano, you can store them in a freezer (-20ºC or better still -80ºC) protected from light for a very long time - decades!
Agreed. In fact lyophilised if light protected they are sufficiently stable that you can store them at room temp for days with no adverse affects or months in the fridge
At lyophilized state and -20ºC and foil-covered, it should be no problem to be stable during your holiday!
Lyophilised primers when stored at -20ºC are often good for up to 18 months, but this may vary depending on supplier.
If you keep your lyophilized primer at -80ºC even for long duration (for years) without any adverse effect
In lypholized stored under perfectly dry conditions, primers can be safely stored at RT or better at -20C for extended period may be upto 5 years. This is most likely the maximum period, before you need to use them.
Primers vials come with label indicating its amount in nM. Just dissolve whole primers in 10 times (of figure indicating its quantity in nM) of 10 mM Tris.cl pH 8.0. Using Tris EDTA BE AVOIDED AS EDTA DUE TO ITS CHELLATING EFFECTON Mg may interfere with normal amp,ifucation.
When I get my primer which comes as lyophilized, I resuspend it to 100 uM/L concentration (stock solution). For example, for an amount of 28.5 nM of lyophilized primer (the amount is provided by the vendor), I add 285 ul water or 10mM Tris.cl (10 time diluted TE) and get 100 uM/L stock solution. To prepare 10 uM/L (or 10 pm/ul) working solution, I take 10 ul stock solution and add to 90 ul water. THIS IS STRAIGHT AND NO FURTHER CALCULATION IS NEEDED.
Hi Pfano
I completely agree with Virash Gupta, this is the best and easier way to get your primers diluted.
Adding to my previous answer, additional care is not repeatedly freeze and thaw your working primer solutions. Just prepare enough vials of working primers (around 200ul each) and use them completely before drawing next vial for use. Prefer storing working solutions at -80C.
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