FastDigest ScaI

Protocol tips

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	ScaI and PvuII pCB568 fragments were obtained by ScaI and PvuII digestion (FastDigest, ThermoFisher) followed by a gel purification of the desired fragment (NucleoSpin Gel and PCR clean-up, Macherey-Nagel).

Protocol tips
ScaI and PvuII pCB568 fragments were obtained by ScaI and PvuII digestion (FastDigest, ThermoFisher) followed by a gel purification of the desired fragment (NucleoSpin Gel and PCR clean-up, Macherey-Nagel).

Publication protocol

"Integrase Substrates Production
Supercoiled plasmids were extracted from E. coli XL1-Blue
using NucleoSpin Plasmid (Macherey-Nagel) or NucleoBond
Xtra Midi (Macherey-Nagel) accordingly to the manufacturer
instructions. Relaxed pCB568 and pCB598 were obtained by
Nt.BspQI digestion (NEB) followed by a column purification
(NucleoSpin Gel and PCR clean-up, Macherey-Nagel). ScaI
and PvuII pCB568 fragments were obtained by ScaI and
PvuII digestion (FastDigest, ThermoFisher) followed by a gel
purification of the desired fragment (NucleoSpin Gel and PCR
clean-up, Macherey-Nagel). Linear pCB598 was obtained by
ScaI digestion (FastDigest, ThermoFisher) followed by a gel purification (NucleoSpin Gel and PCR clean-up, Macherey-Nagel).
A 2,106-bp fragment of pCB568 was amplified by Phusion
Polymerase (ThermoFisher) with the primers pUC1481-1503
and P30-REV followed by column purification (NucleoSpin
Gel and PCR clean-up, Macherey-Nagel). The various fragments
of 800 bp were amplified from the appropriate plasmid by
Phusion Polymerase (ThermoFisher) with the primers
pUC195-217 and pZE21_rev followed by column purification
(NucleoSpin Gel and PCR clean-up, Macherey-Nagel)."

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Manufacturer protocol

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