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The best way of doing the ligation is to set up two digests in two tubes: Digest 1: Cut vector with PstI and NdeI and isolate 4627 bp band from gel. Digest 2: Cut vector with PstI and XhoI and
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The concentration of the Guanidine HCl must be below 250 mM for the ULP1 digestion to be effective. 2. Add ULP1 and incubate with stirring at 4 °C. 3.
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