Your primers will last longer in buffered solution than water. I always suspend my master stock of primers at 100uM in 10mM Tris-HCL, pH 8.5. I then dilute these to 10uM stocks that I use in downstream applications like PCR.
Your primers will last longer in buffered solution than water. I always suspend my master stock of primers at 100uM in 10mM Tris-HCL, pH 8.5. I then dilute these to 10uM stocks that I use in downstream applications like PCR.
If water is used I think- The pH of water can be low (especially if it is DEPC treated) and this will be damaging to primer (DNA) over time.
This is what I found:
To resuspend/ reconstitute PRIMERS:
Use a buffered solution at neutral pH to protect from acid hydrolysis. EDTA (1 mM) in the master stock is also a good idea to protect against DNases and when you dilute the primers for working stocks, the EDTA will be sufficiently dilute that it will not interfere with Taq activity.
I have been using normal milliQ water and never had any problem. 10 mM Tris/HCl pH 8.5 might be better though.
All nucleic acids should be dissolved in pH-controlled solutions as CO2 from the air will lower pH of pure water and hydrolize nucleic acids. I use generally TE buffer 10mM Tris, 1mM EDTA, pH8.5). The EDTA contration is low enough. Consequently it will not interfer with enzyme reactions after dilution for the enzyme treatment. It is also strongly recommended to aliquot primers to avoid frequent thawing and re-freezing. I use batches not used more than 5 times.
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