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URL: http://www.addgene.org/52672
Proper Citation: RRID:Addgene_52672
Insert Name: PSD-95
Organism: Rattus norvegicus
Bacterial Resistance: Kanamycin
Vector Backbone Description: Backbone Marker:Evrogen; Backbone Size:4733; Vector Backbone:pTagRFP; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: HindII and EcoRI were used to cut PSD-95 from PSD-95-pGW1 (received from David Bredt) and ligate it into pTagRFP-N. PSD-95 tyrosine 12 was mutated to glutamate (Y12E) by extending a complementary set of primers (PSD95Y12E-forward: 5’-GAC AAC CAA GAA AGA GCG CTA CCA AGA TGA AGA CAC G-3’, PSD95Y12E-reverse: 5'-CGT GTC TTC ATC TTG GTA GCG CTC TTT CTT GGT TGT C-3') along the whole template plasmid using Q5 high-fidelity DNA polymerase (NEB). DpnI was subsequently used to digest non-mutated, methylated template DNA prior to transformation.
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Source: Addgene
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