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How can I facilitate the amplification of templates with hairpin-loop structures or high CG-content?

April 26, 2017amplification facilitate high structures Templates

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How can I facilitate the amplification of templates with hairpin-loop structures or high CG-content?

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You can try 5-10 % DMSO, up tp 10 % glycerol, 1-2 % formamide or combinations of these. Note: The use of co-solvents will lower the optimal annealing temperatures of your primers (e.g. 5.5-6 °C in 10 % DMSO). You could also try 7-deaza-dGTP in conjuction with normal dGTP in order to destabilize difficult structures. Note: 7-deaza-dGTP attenuates the signal of ethidium bromide staining.