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

DNA constructs
WTFUS and mFUS (P525L, R524S, R522G and R521G) constructs encoding HA-tagged human FUS at the N-terminus were a gift from Dr Dorothee Dormann, Ludwig Maximilians Universität München, Munich. EGFP-Q74 vector was a gift from David Rubinsztein (Addgene plasmid # 40262, Cambridge, MA, USA).49 mCherry-EGFP-LC3 vector was a gift from Jayanta Debnath (Addgene plasmid # 22418).50 Dsred-LC3 vector was a kind gift from Dr Yung-Feng Liao, Institute of Cellular and Organismic, Academia Sinica, Taiwan.51 Myc-DFCP1 and EGFP-DFCP1 vectors were kind gift from Dr Nicholas Ktistakis, Babraham Institute, United Kingdom. CFP-Rab1a vector was a kind gift from Professor Harald H. Sitte, Medical University Vienna. Mutant Rab1, S25N and Q70L, were generated by using Q5 Site-Directed Mutagenesis kit (New England Biolabs, Ipswich, MA, USA). p62-GFP vector was a kind gift from Professor Guanghui Wang, Soochow University College of Pharmaceutical Sciences, Suzhou. pcDNA-LAMP2C vector was a kind gift from Professor Tomohiro Kabuta, National Center of Neurology and Psychiatry; Kodaira, Tokyo, Japan.

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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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