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HotStarTaq PCR Handbook

ution enables amplification of a reaction which previously failed.
Case B
: Q-Solution increases PCR specificity in certain primertemplate systems.
Case C
: Q-Solution has no effect on PCR performance.
Case D
: Q-Solution causes reduced efficiency or failure of a previously successful amplification reaction. In this case, addition of Q-Solution disturbs the previously optimal primertemplate annealing. Therefore, when using Q-Solution for the first time for a particular primertemplate system, always perform reactions in parallel with and without Q-Solution.



Notes:
HotStarTaq DNA Polymerase requires an activation step of 15 min at 95C (see step 6 of this protocol).
When using Q-Solution for the first time in a particular primertemplate system, it is important to perform parallel amplification reactions with and without Q-Solution.
Set up all reaction mixtures in an area separate from that used for DNA preparation or PCR product analysis.
Use disposable tips containing hydrophobic filters to minimize cross-contamination.
If required, prepare a dNTP mix containing 10 mM of each dNTP. Store this mix in aliquots at 20C.

1. Thaw 10x PCR Buffer, dNTP mix, primer solutions, and Q-Solution.
It is important to mix the solutions completely before use to avoid localized concentrations of salts. When using Q-Solution, additional MgCl2 is not usually
required.

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Source:


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