Recently I've been trying to amplify 4kb using the phusion polymerase, but I have doubts with the annealing temperature, can somebody recomend an article talking about that. and give me some advice?
but can you give me some advices, is the first time in use of this polimerase and i want to make all the calculations for the PCR conditions and concentrations thx!
but you controle the Mg concentration only decreasing the Buffer HF 5X concentration or you have different technique :)
may be this article could help you:
Breslauer, K J., Frank, R., Blöcker, H., Marky, L A., (1986) Predicting DNA duplex stability from the base sequence. PNAS 83 (11), 3746-50
Antonio Gracias! lo leere de inmediato , Brad i have a few questions and a image of the gradient temp if u can help me thanks
Hi! Can you please give me the name of your enzyme? I´m trying to amplify 2400 bp but I can´t. I´m using High Fidelity and Pfu. Please, help me! Thanks :)
claro, Phusion Polymerase i sent you the link of the catalogue http://www.fermentas.com/en/products/all/pcr-qpcr-rt-pcr/high-fidelity-pcr/f530-phusion-high-fidelity-dna-polymerase
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