Q5® Site-Directed Mutagenesis Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Only use 1 μl of PCR product in the KLD reaction. Carrying too much PCR product forward can decrease transformation efficiency. - Only use 5 μl of the KLD reaction in the transformation. If more KLD reaction is added, a buffer exchange step, such as PCR purification, should be included prior to transformation.

Protocol tips
- Only use 1 μl of PCR product in the KLD reaction. Carrying too much PCR product forward can decrease transformation efficiency. - Only use 5 μl of the KLD reaction in the transformation. If more KLD reaction is added, a buffer exchange step, such as PCR purification, should be included prior to transformation.

Publication protocol

All Epac1 mutant constructs were generated using either GENEART® Site-Directed Mutagenesis System from Invitrogen (Carlsbad, CA, USA) or Q5® Site-Directed Mutagenesis Kit from NEB and validated by DNA sequencing. Importin β1 si-RNA (FlexiTube Mm_Kpnb1_5) and (FlexiTube Hs_KPNB1_1), and control si-RNA (AllStars Neg. Control) were purchased from Qiagen (Cambridge, MA, USA).

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Discussion

Papers

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Manufacturer protocol

Download the product protocol from New England BioLabs for Q5® Site-Directed Mutagenesis Kit below.

Download manufacturer protocol

Videos

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