I try to cut them with these enzyme in separat reaction, to linearise the plasmid but it did not work, Although I incubate them for 2hours at 37 heat block.
Touraj Miandoabi
Incubate at 37°C for 5-15 minutes as BamHI is Time-Saver qualified. Add 1.0 µl (or 10 units) of BclI Incubate at 50°C for 5-15 minutes as BclI is Time-Saver qualified. I hope you get the good result. good luck
Hi,
BClI is typically used at 50-55 °c, but is 50% active at 37°c.
Set up reaction as follows: COMPONENT 50 µl REACTION
pCAMBIA 2300Plasmid 1 µg 10X Buffer 5 µl (1X) BamHI 1.0 µl (or 10 units) Nuclease-free Water to 50 µl
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Mona Albloushi
Hi Touraj
Thanks for your advice. I dillued my plasmid and start the reaction.
Many Thanks
Mona
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