( ...)QIAGEN Multiplex PCR Handbook

3. Handling and storage of primers

Primer quality is crucial for successful multiplex PCR. Problems encountered with multiplex PCR are frequently due to incorrect concentrations of primers being used. If you observe large differences in yield of different amplification products in a multiplex PCR, check that all primers were used at the correct concentration. For optimal results, we recommend only combining purified primers of comparable quality.

Table 11. General guidelines for handling and storage of PCR primers
Primer storage buffer: Lyophilized primers should be dissolved in a small volume of low salt buffer to make a concentrated stock solution. We recommend using TE (10 mM TrisCl, 1 mM EDTA, pH 8.0) for standard primers and primers labeled with most fluorescent dyes. However, primers labeled with fluorescent dyes such as Cy3, Cy3.5, Cy5, and Cy5.5, should be stored in TE, pH 7.0 since they tend to degrade at higher pH. Dissolving primers: Before opening tubes containing lyophilized primers, spin tubes briefly to collect all material at the bottom of the tube. To dissolve the primer, add the required volume of TE, mix, and leave for 20 minutes to let the primer completely dissolve. We do not recommend dissolving primers in water. Primers are less stable in water than TE and some primers may not dissolve easily in water. Storage: Primers should be stored in TE in small aliquots at 20C. Unmodified primers are stable under these conditions for at le
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