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
The wild-type human FGF1 coding region was subcloned into a constitutive mammalian expression vector (pcDNA3.1/V5-His, Invitrogen, Waltham, MA, USA) allowing the fusion with a C-terminal V5-His tag (pcDNA3.1-FGF1WT). Mutant FGF1 expression vectors were generated using the QuickChange II XL Site-Directed Mutagenesis Kit (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s protocol. We used the pcDNA3.1-FGF1WT vector as a DNA template to generate pcDNA3.1-FGF1S130A, pcDNA3.1-FGF1S130D, pcDNA3.1-FGF1K132E with specific primers. These vectors allow the expression of C-terminal V5-His tagged wild-type or mutant FGF1 (aa 15 to 154).
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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 SH-SY5Y FGF1 using QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn from Agilent Technologies.
Paper title
FGF1 protects neuroblastoma SH-SY5Y cells from p53-dependent apoptosis through an intracrine pathway regulated by FGF1 phosphorylation
Manufacturer protocol
Download the product protocol from Agilent Technologies for QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn below.
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