Optimal reaction conditions for double digestion with KpnI and XhoI?

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Jorge Barros-Velázquez

Hi Surya:

I strongly recommend you to use the "double calculator" tool from ThermoFisher:

https://www.thermofisher.com/us/en/home/brands/thermo-scientific/molecular-biology/thermo-scientific-restriction-modifying-enzymes/restriction-enzymes-thermo-scientific/double-digest-calculator-thermo-scientific.html

In your particular case you would need to perform it at 37oC in two steps with two different buffers:

Best regards

Jorge

Chunhong Chen

Hi, Surya,

If you buy KpnI-HF and Xho I from NEB, you can set up double digestion in CutSmart Buffer. Good luck!

Chunhong

Qing Zhao

KpnI and XhoI, from which company? I seem to remember the NEB's two products digestion temperature is not the same?

Surya Prasad Tiwari

Thanks everyone for your nice suggestions & comments

Qing Zhao: I'd used KpnI and XhoI, which were from Thermo Fisher Scientific.