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Species: Homo sapiens
Genetic Insert: Desmoplakin
Vector Backbone Description: Backbone Size:5000; Vector Backbone:pCMV; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32398 Copy
Species: Homo sapiens
Genetic Insert: ATM
Vector Backbone Description: Backbone Marker:invitrogen; Backbone Size:5428; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32300 Copy
Species: Homo sapiens
Genetic Insert: SMC1A
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pCDNA3 cMyc; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32371 Copy
Species: Homo sapiens
Genetic Insert: SMC1A
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pCDNA3 cMyc; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32372 Copy
Species: Homo sapiens
Genetic Insert: SMC1A
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pCDNA3 cMyc; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32363 Copy
Species: Homo sapiens
Genetic Insert: GSK3B
Vector Backbone Description: Backbone Marker:Sabatini Lab; Backbone Size:7000; Vector Backbone:pLKO.1; Vector Types:Mammalian Expression, Lentiviral, RNAi; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32496 Copy
Species: Homo sapiens
Genetic Insert: Ataxin-1
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pEGFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_32492 Copy
Species: Homo sapiens
Genetic Insert: HSF1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Please note that mutation P448L in HSF1 was found during Addgene's quality control. This plasmid should function as reported in the associated publication.
Proper citation: RRID:Addgene_32537 Copy
Species: Homo sapiens
Genetic Insert: HSF1
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4729; Vector Backbone:pEGFP-N3; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments: Please note that mutation P448L in HSF1 was found during Addgene's quality control. This plasmid should function as reported in the associated publication.
Proper citation: RRID:Addgene_32538 Copy
Species: Homo sapiens
Genetic Insert: PSAML141F,Y115F-GlyR
Vector Backbone Description: Backbone Size:5100; Vector Backbone:AAV2; Vector Types:AAV, Cre/Lox, Adeno-associated virus, FLEX switch; Bacterial Resistance:Ampicillin
References:
Comments: Comments:
These rAAV vectors are prone to recombination. This is a well known issue with these rAAV vectors and is due to the inverted terminal repeats (ITRs) required for rAAV production. To minimize recombination, we propagate these plasmids in NEB Stable cells. Also, to minimize recombination, cells should be cultured at 30 C.
Note that these cultures will grow slowly (20 h for minipreps). Better yields and culture times are obtained with 2xYT as the media. This is strongly recommended.
Because recombination may still happen, we do a panel of restriction digestions to assess whether the ITRs are in tact. Separate digestions with Sma1 and Msc1 should be performed. The expected patterns can be calculated from the attached sequence.
In Magnus et al, the PSAMY115F,L141F-GlyR channel was emphasized for silencing because this channel has the lowest acetylcholine responsiveness. However, PSAML141F-GlyR constructs, lacking the Y115F mutation, already have low ACh potency. The Y115F mutation also slightly increases the EC50 for the activator ligand, PSEM89S. Since PSAML141F-GlyR constructs already have very low acetylcholine potency, in many cases it may not be worth the slight reduction in PSEM potency. We have made both constructs available for researchers who may find that they have different requirements with respect to acetylcholine responsiveness.
Proper citation: RRID:Addgene_32481 Copy
Species: Homo sapiens
Genetic Insert: Protein tyrosine phosphatase receptor-type delta G1707R
Vector Backbone Description: Backbone Marker:System Biosciences ; Backbone Size:6606; Vector Backbone:pCDF1-MCS2-EF1-Puro; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32526 Copy
Species: Homo sapiens
Genetic Insert: Protein tyrosine phosphatase receptor-type delta P1690F
Vector Backbone Description: Backbone Marker:System Biosciences ; Backbone Size:6606; Vector Backbone:pCDF1-MCS2-EF1-Puro; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32525 Copy
Species: Homo sapiens
Genetic Insert: PSAMQ79G,Q139G-5HT3 HC
Vector Backbone Description: Backbone Size:7200; Vector Backbone:pCAGGS- IRES-GFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32474 Copy
Species: Homo sapiens
Genetic Insert: PSAMQ79G,Q139G-5HT3HC
Vector Backbone Description: Backbone Size:5100; Vector Backbone:AAV2; Vector Types:AAV, Cre/Lox, Adeno-associated virus, FLEX switch; Bacterial Resistance:Ampicillin
References:
Comments: These rAAV vectors are prone to recombination. This is a well known issue with these rAAV vectors and is due to the inverted terminal repeats (ITRs) required for rAAV production. To minimize recombination, we propagate these plasmids in NEB Stable cells. Also, to minimize recombination, cells should be cultured at 30 C.
Note that these cultures will grow slowly (20 h for minipreps). Better yields and culture times are obtained with 2xYT as the media. This is strongly recommended.
Because recombination may still happen, we do a panel of restriction digestions to assess whether the ITRs are in tact. Separate digestions with Sma1 and Msc1 should be performed. The expected patterns can be calculated from the attached sequence.
Proper citation: RRID:Addgene_32475 Copy
Species: Homo sapiens
Genetic Insert: PSAM Y115F,L141F-5HT3 HC
Vector Backbone Description: Backbone Size:7200; Vector Backbone:pCAGGS- IRES-GFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32476 Copy
Species: Homo sapiens
Genetic Insert: NRBP
Vector Backbone Description: Backbone Marker:Agilent; Backbone Size:4100; Vector Backbone:pSG5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32504 Copy
Species: Homo sapiens
Genetic Insert: CIB1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_32505 Copy
Species: Homo sapiens
Genetic Insert: p66shc
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5500; Vector Backbone:pcDNA3.1 His; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: This plasmid is a corrected version of pcDNA3.1his p66shc (Addgene plasmid 10972 http://www.addgene.org/10972). D2N and S60P mutations present in the original sample have been corrected in this plasmid.
Proper citation: RRID:Addgene_32574 Copy
Species: Homo sapiens
Genetic Insert: RAP1
Vector Backbone Description: Vector Backbone:pAXEF-1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin and Tetracycline
References:
Comments: Q63 changed to E to give phenotype
Proper citation: RRID:Addgene_32698 Copy
Species: Homo sapiens
Genetic Insert: GRK4
Vector Backbone Description: Backbone Size:4754; Vector Backbone:pRK5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Contains the longest splice variant of GRKa. The insert does not contain the 3'UTR.
Proper citation: RRID:Addgene_32690 Copy
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