GeneArt™ Site-Directed Mutagenesis System

Site Directed Mutagenesis (SDM) Human - Point mutation LNCaP RasGRP2

Experiment
Site Directed Mutagenesis (SDM) Human - Point mutation LNCaP RasGRP2
Product
GeneArt™ Site-Directed Mutagenesis System from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
- Use 20–25 ng of the target plasmid per 50 µL of mutagenesis reaction as a starting point.
Protocol tips
- Create a fresh dilution of 25X SAM from the kit-supplied 200X SAM in sterile, distilled water each time you perform the mutagenesis procedure
- 25X SAM is not stable, and loses activity within a few hours after
preparation

Publication protocol

Site-directed Mutagenesis of the C1 Domain and the Full-length RasGRP2
Point mutations of the amino acid residues were introduced using the GeneTailor® and GeneArt® site-directed mutagenesis system (Invitrogen) according to the manufacturer's instructions. To generate the C1 domain and full-length mutants of RasGRP2, the above mentioned wild-type C1 and full-length constructs in pEGFP-C3 and pGEX-2T were used. Single mutations (S8Y) were introduced in one step, and double (S8Y/A19G), triple (N7T/S8Y/A19G and S8Y/A19G/I21L), and quadruple mutants (N7T/S8Y/A19G/I21L) were generated in a stepwise fashion using single and double mutants as templates. The presence of mutations was verified by DNA sequencing (DNA Minicore).

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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 LNCaP RasGRP2 using GeneArt™ Site-Directed Mutagenesis System from Thermo Fisher Scientific.

Paper title
Structural Basis for the Failure of the C1 Domain of Ras Guanine Nucleotide Releasing Protein 2 (RasGRP2) to Bind Phorbol Ester with High Affinity
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for GeneArt™ Site-Directed Mutagenesis System below.

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