( ...)HotStarTaq PCR Handbook

Little or no product 1. HotStarTaq DNA Polymerase not activated Check whether PCR was started with an initial incubation step at 95C for 15 min. 2. Pipetting error or missing reagent Repeat the PCR. Check the concentrations and storage conditions of reagents, including primers and dNTP mix. In the case of HotStarTaq Master Mix, ensure that a 1:1 ratio of HotStarTaq Master Mix to primertemplate solution is maintained. 3. PCR cycling conditions are not optimal Using the same cycling conditions, repeat the PCR using Q-Solution. 4. Primer concentration not optimal
or primers degraded Repeat the PCR with different primer concen trations from 0.10.5 M of each primer (in 0.1-M steps). In particular, when performing highly sensitive PCR, check for possible degradation of the primers on a denaturing polyacrylamide gel. 5. Problems with starting template Check the concentration, storage conditions, and quality of the starting template (see appendix). If necessary, make new serial dilutions of template nucleic acid from stock solutions. Repeat the PCR using the new dilutions.
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