Site directed mutagenesis reagent

Manufactured by Agilent Technologies

The Site-directed mutagenesis reagent is a laboratory tool used to introduce specific genetic mutations into DNA sequences. It provides the necessary components to carry out this process, which is a widely used technique in molecular biology and genetics research.

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Lab products found in correlation

Cycloheximide (chx), by Santa Cruz Biotechnology (1 mentions) Recombinant glua1, by OriGene (1 mentions) Poly d lysine, by Santa Cruz Biotechnology (1 mentions) Anti pan 14 3 3, by Merck Group (1 mentions) Anti glua1, by Abcam (1 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions) Anti map2, by Thermo Fisher Scientific (1 mentions) Anti α tubulin, by Cell Signaling Technology (1 mentions) Protein a g bead, by Santa Cruz Biotechnology (1 mentions) Anti gapdh, by GenScript (1 mentions) Anti ha, by GenScript (1 mentions) Pgl3 control vector, by Promega (1 mentions)

2 protocols using site directed mutagenesis reagent

Epilepsy-Associated Nedd4-2 Mutant Characterization

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Dimethyl sulfoxide (DMSO) was from Fisher Scientific. AMPA was from Cayman Chemical and NBQX was from Alomone Labs. Recombinant GluA1 and 14-3-3ε were from Origene. Recombinant Nedd4-2 was from Abnova. R18 was from Sigma-Aldrich. Cycloheximide, poly-D-lysine and Protein A/G beads were from Santa Cruz Biotechnology. The antibodies used in this study were purchased from Santa Cruz Biotechnology (anti-α-Tubulin), Cell Signaling (anti-Nedd4-2, anti-pan-14-3-3, anti-N-cadherin and anti-Ubiquitin), Millipore (anti-GluA1), Abcam (anti-MAP2), Thermo Scientific (anti-HA) and GenScript Corporation (anti-Gapdh). The epilepsy-associated mutations were generated using site-directed mutagenesis reagent (Agilent) to introduce mutations into pCI-HA-Nedd4-2 [15 (link)]. The primers used are as below.
S233L: 5’-GGACGTGTCCTCGGAGTTGGACAATAACATCAGAC-3’,
5’-GTCTGATGTTATTGTCCAACTCCGAGGACACGTCC-3’;
E271A: 5’- GGGCGGGGATGTCCCCGCGCCTTGGGAGACCATTTC-3’,
5’- GAAATGGTCTCCCAAGGCGCGGGGACATCCCCGCCC-3’;
H515P: 5’- CGTTTGAAATTTCCAGTACCTATGCGGTCAAAGACATC-3’,
5’- GATGTCTTTGACCGCATAGGTACTGGAAATTTCAAACG-3’.

Zhu J., Lee K.Y., Jewett K.A., Man H.Y., Chung H.J, & Tsai N.P. (2017). Epilepsy-associated gene Nedd4-2 mediates neuronal activity and seizure susceptibility through AMPA receptors. PLoS Genetics, 13(2), e1006634.

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Cloning and Mutagenesis of PIM2 3'-UTR

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Sequences of putative site of PIM2′s 3′-UTR (untranslated region) that harbor the miR-135-5p were amplified through PCR with random primers and cloned into the XbaI site of a pGL3-Control vector (Promega, Madison, WI). Mutant constructs of the same miRNA seed sequences were generated using specific primers and the Site-Directed Mutagenesis reagent (Agilent Technologies, Santa Clara, CA) according to the manufacturer’s instructions. The mutation was verified by direct sequencing.

, & Zhang H. (2017). Upregulation of PIM2 by Underexpression of MicroRNA-135-5p Improves Survival Rates of Skin Allografts by Suppressing Apoptosis of Fibroblast Cells. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 23, 107-113.

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