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

Constructs
FLAG/HIS tagged APRIN was cloned by polymerase chain reaction (PCR) amplification of pCMV SPORT plasmid APRIN-myc-his (gift from Peter Geck) into pFastBAC1-52b-GST using the primers cited in Table . pEGFP-APRIN was obtained after the insertion of EcoR1/BamH1 PCR products into pEGFP-C1 (Clontech). APRIN deletion constructs were obtained by mutagenesis using the Q5 Site-Directed Mutagenesis Kit (NEB) and primers cited in Table . APRIN fragments were cloned by PCR amplification of pCMV SPORT plasmid APRIN-myc-his (gift from Peter Geck) into pEGFP-C1 using primers cited in Table . mCherry-PCNA-19-NLS-4 was purchased from Addgene.

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Reviews

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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