Q5® Site-Directed Mutagenesis Kit

Site Directed Mutagenesis (SDM) Human - Point mutation U-87MG Rictor

Experiment
Site Directed Mutagenesis (SDM) Human - Point mutation U-87MG Rictor
Product
Q5® Site-Directed Mutagenesis Kit from New England BioLabs
Manufacturer
New England BioLabs

Protocol tips

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

To generate the deletion construct of Rictor, we separated the previously reported multiacetylation domain of Rictor into four parts (Δ1080–1089, Δ1089–1098, Δ1098–1110, and Δ1110–1128), according to predicted acetylated lysines within Rictor in silico (Fig. S2), and we performed the PCR-based construction of deletion. To create the 3KR and 3KQ mutants using the Myc-Rictor DNA plasmid, we replaced three predicted acetylation sites—K1116, K1119, and K1125—with R for the KR mutant and Q for the KQ mutant. We carried out site-directed mutagenesis using the Quick Change Kit (Stratagene) or Q5 Site-Directed Mutagenesis Kit (New England Biolabs).

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Site Directed Mutagenesis (SDM) Human - Point mutation U-87MG Rictor using Q5® Site-Directed Mutagenesis Kit from New England BioLabs.

Paper title
Glucose-dependent acetylation of Rictor promotes targeted cancer therapy resistance
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Manufacturer protocol

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

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