Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
- XL10-Gold cells are resistant to tetracycline and chloramphenicol. If the mutagenized plasmid contains only the tetR or camR resistance marker, an alternative strain of competent cells must be used |
- Use only the Dpn I enzyme provided; do not substitute with an enzyme from another source. |
|
Upstream tips |
- XL10-Gold cells are resistant to tetracycline and chloramphenicol. If the mutagenized plasmid contains only the tetR or camR resistance marker, an alternative strain of competent cells must be used |
Protocol tips |
- Use only the Dpn I enzyme provided; do not substitute with an enzyme from another source. |
Publication protocol
Site-directed Mutagenesis
PKC phosphorylation sites were identified using LC-MS/MS on human CELF1 protein expressed in 293T cells. The predicted sites of Ser (28, 52, 178, 179, 241, 300, 302) were mutated to Ala and Thr-173 to Val using site-directed mutagenesis kit (Stratagene 200516) to generate the CELF1M8 construct in p3×FLAG vector.
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Papers
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Paper title
Reactivation of Fetal Splicing Programs in Diabetic Hearts Is Mediated by Protein Kinase C Signaling
Manufacturer protocol
Download the product protocol from Agilent Technologies for QuikChange Site-Directed Mutagenesis Kit, 10 Rxn below.
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