Stratagene quikchange method

Manufactured by Agilent Technologies

The Stratagene QuikChange method is a site-directed mutagenesis technique used to introduce specific mutations into double-stranded plasmid DNA. The method allows for the rapid and efficient mutation of one or more bases in a DNA sequence.

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QuikChange (236 citations) QuikChange method (57 citations) Stratagene QuikChange (5 citations)

4 protocols using stratagene quikchange method

Site-directed Mutagenesis of GPR84-G i2

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The Stratagene QuikChange method
(Stratagene, Agilent Technologies, Santa Clara, CA) was used to introduce
alterations into FLAG-human GPR84-Gα_i2 or FLAG-mouse
GPR84-Gα_i2. Primers utilized for mutagenesis were
provided by MWG Operon (Acton, UK). Template DNA was digested with
DpnI to leave only the newly synthesized mutated plasmid, and sequencing
was carried out to confirm the introduction of the alterations.

Jenkins L., Marsango S., Mancini S., Mahmud Z.A., Morrison A., McElroy S.P., Bennett K.A., Barnes M., Tobin A.B., Tikhonova I.G, & Milligan G. (2021). Discovery and Characterization of Novel Antagonists of the Proinflammatory Orphan Receptor GPR84. ACS Pharmacology & Translational Science, 4(5), 1598-1613.

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Generation and Characterization of Mutant GPR35 Constructs

Cited 1 time

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Generation of FLAG-hGPR35a-eYFP, hGPR35a-HA, FLAG-mGPR35-eYFP, and mGPR35-HA have been described previously (35 (link), 36 (link)). The Stratagene Quik Change method (Stratagene, Agilent Technologies) was used to introduce alterations into each of the above constructs to produce both point mutants and phospho-deficient variants. Primers utilised for mutagenesis were from MWG Operon. Sequencing was carried out to confirm the introduction of the alterations.

Ganguly A., Quon T., Jenkins L., Joseph B., Al-awar R., Chevigne A., Tobin A.B., Uehling D.E., Hoffmann C., Drube J, & Milligan G. (2023). G protein–receptor kinases 5/6 are the key regulators of G protein–coupled receptor 35–arrestin interactions. The Journal of Biological Chemistry, 299(10), 105218.

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Mutagenesis of GPR84 Receptor

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The Stratagene QuikChange method (Stratagene, Agilent Technologies) was used to introduce alterations into FLAG-human GPR84-eYFP or FLAG-human GPR84-Gα_i2. Primers utilized for mutagenesis were provided by MWG Operon. Template DNA was digested with DpnI to leave only the newly synthesized mutated plasmid, and sequencing was carried out to confirm the introduction of the alterations.

Marsango S., Ward R.J., Jenkins L., Butcher A.J., Al Mahmud Z., Dwomoh L., Nagel F., Schulz S., Tikhonova I.G., Tobin A.B, & Milligan G. (2022). Selective phosphorylation of threonine residues defines GPR84–arrestin interactions of biased ligands. The Journal of Biological Chemistry, 298(5), 101932.

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Introducing Mutations in VSV-SNAP-hD3

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The Stratagene QuikChange method (Stratagene, Agilent Technologies, Santa Clara, CA) was used to introduce alterations into VSV-SNAP-hD₃. Primers utilized for mutagenesis were provided by MWG Operon (Acton, UK). Template DNA was digested with DpnI to leave only the newly synthesized mutated plasmid, and sequencing was carried out to confirm the introduction of the alterations.

Marsango S., Caltabiano G., Pou C., Varela Liste M.J, & Milligan G. (2015). Analysis of Human Dopamine D3 Receptor Quaternary Structure. The Journal of Biological Chemistry, 290(24), 15146-15162.

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