rp11-265b8 Search Results


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  • 99
    Vector Laboratories avidin d texas red
    Avidin D Texas Red, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 268 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    bg01v  (ATCC)
    93
    ATCC bg01v
    Bg01v, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Roche digoxigenin
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Digoxigenin, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 10258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories biotinylated anti avidin d
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Biotinylated Anti Avidin D, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 98/100, based on 365 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    GE Healthcare cy5 ap3 dutp
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Cy5 Ap3 Dutp, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 102 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Roche expand long template pcr kit
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Expand Long Template Pcr Kit, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 472 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TaKaRa pdsred express n1 plasmid
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Pdsred Express N1 Plasmid, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Roche pcr amplification
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Pcr Amplification, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 13428 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen long range pcr kit
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Long Range Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 281 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    The Jackson Laboratory cat lutz
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Cat Lutz, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 89/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    TaKaRa cmv promoter
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Cmv Promoter, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 2526 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Gene Bridges Inc redet recombination
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Redet Recombination, supplied by Gene Bridges Inc, used in various techniques. Bioz Stars score: 92/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Promega pgl4 10 luc2
    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-labelled</t> RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.
    Pgl4 10 Luc2, supplied by Promega, used in various techniques. Bioz Stars score: 89/100, based on 388 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore hybridisation buffer
    Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three <t>hybridisation</t> signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.
    Hybridisation Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 324 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Roche cot 1 human dna
    Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three <t>hybridisation</t> signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.
    Cot 1 Human Dna, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 136 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa dsred express gene
    Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three <t>hybridisation</t> signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.
    Dsred Express Gene, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human fibroblast feeders ws1
    Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three <t>hybridisation</t> signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.
    Human Fibroblast Feeders Ws1, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore anti rabbit fitc
    Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three <t>hybridisation</t> signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.
    Anti Rabbit Fitc, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 332 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore mouse anti digoxigenin antibody
    Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and <t>digoxigenin-</t> labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three hybridisation signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.
    Mouse Anti Digoxigenin Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega pgl3 basic vector
    Regulatory effects of a putative Oct-1 binding site located within conserved region 1. (A) Two mutant constructs were generated by site directed mutagenesis within a putative Oct-1 binding site within the identified 17 bp nucleotide sequence. MUT 1 contains one additional extra base in the middle of the sequence and MUT 2 contains two bases substitution at the start of the sequence. (B) MUT 1 and MUT 2 were generated in the <t>pGL3-Basic::5023</t> construct. Luciferase expression of the mutant constructs was analyzed following co-transfection of BE(2)-M17 cells with pGL3-Basic derivatives and Oct-1 transcription factor-encoding plasmid pCMV-AC::Oct-1. The overall levels of expression were then compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p
    Pgl3 Basic Vector, supplied by Promega, used in various techniques. Bioz Stars score: 95/100, based on 27770 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Syngene genetools software
    Regulatory effects of a putative Oct-1 binding site located within conserved region 1. (A) Two mutant constructs were generated by site directed mutagenesis within a putative Oct-1 binding site within the identified 17 bp nucleotide sequence. MUT 1 contains one additional extra base in the middle of the sequence and MUT 2 contains two bases substitution at the start of the sequence. (B) MUT 1 and MUT 2 were generated in the <t>pGL3-Basic::5023</t> construct. Luciferase expression of the mutant constructs was analyzed following co-transfection of BE(2)-M17 cells with pGL3-Basic derivatives and Oct-1 transcription factor-encoding plasmid pCMV-AC::Oct-1. The overall levels of expression were then compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p
    Genetools Software, supplied by Syngene, used in various techniques. Bioz Stars score: 92/100, based on 3910 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy mini kit
    Regulatory effects of a putative Oct-1 binding site located within conserved region 1. (A) Two mutant constructs were generated by site directed mutagenesis within a putative Oct-1 binding site within the identified 17 bp nucleotide sequence. MUT 1 contains one additional extra base in the middle of the sequence and MUT 2 contains two bases substitution at the start of the sequence. (B) MUT 1 and MUT 2 were generated in the <t>pGL3-Basic::5023</t> construct. Luciferase expression of the mutant constructs was analyzed following co-transfection of BE(2)-M17 cells with pGL3-Basic derivatives and Oct-1 transcription factor-encoding plasmid pCMV-AC::Oct-1. The overall levels of expression were then compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p
    Rneasy Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 343576 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs pbr322 plasmid
    Regulatory effects of a putative Oct-1 binding site located within conserved region 1. (A) Two mutant constructs were generated by site directed mutagenesis within a putative Oct-1 binding site within the identified 17 bp nucleotide sequence. MUT 1 contains one additional extra base in the middle of the sequence and MUT 2 contains two bases substitution at the start of the sequence. (B) MUT 1 and MUT 2 were generated in the <t>pGL3-Basic::5023</t> construct. Luciferase expression of the mutant constructs was analyzed following co-transfection of BE(2)-M17 cells with pGL3-Basic derivatives and Oct-1 transcription factor-encoding plasmid pCMV-AC::Oct-1. The overall levels of expression were then compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p
    Pbr322 Plasmid, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 89/100, based on 212 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher salmon sperm dna
    Regulatory effects of a putative Oct-1 binding site located within conserved region 1. (A) Two mutant constructs were generated by site directed mutagenesis within a putative Oct-1 binding site within the identified 17 bp nucleotide sequence. MUT 1 contains one additional extra base in the middle of the sequence and MUT 2 contains two bases substitution at the start of the sequence. (B) MUT 1 and MUT 2 were generated in the <t>pGL3-Basic::5023</t> construct. Luciferase expression of the mutant constructs was analyzed following co-transfection of BE(2)-M17 cells with pGL3-Basic derivatives and Oct-1 transcription factor-encoding plasmid pCMV-AC::Oct-1. The overall levels of expression were then compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p
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    Thermo Fisher 13x human cot 1 dna
    Regulatory effects of a putative Oct-1 binding site located within conserved region 1. (A) Two mutant constructs were generated by site directed mutagenesis within a putative Oct-1 binding site within the identified 17 bp nucleotide sequence. MUT 1 contains one additional extra base in the middle of the sequence and MUT 2 contains two bases substitution at the start of the sequence. (B) MUT 1 and MUT 2 were generated in the <t>pGL3-Basic::5023</t> construct. Luciferase expression of the mutant constructs was analyzed following co-transfection of BE(2)-M17 cells with pGL3-Basic derivatives and Oct-1 transcription factor-encoding plasmid pCMV-AC::Oct-1. The overall levels of expression were then compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p
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    Image Search Results


    Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin-labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.

    Journal: Disease Models & Mechanisms

    Article Title: A novel GAA-repeat-expansion-based mouse model of Friedreich’s ataxia

    doi: 10.1242/dmm.018952

    Figure Lengend Snippet: Transgene copy number. (A) Two TaqMan copy-number assays were applied: Hs05092416-cn assay (ATX 416; represented in black) was designed to amplify a 106-bp fragment of FXN within intron 3, and Hs02407730-cn assay (ATX 730; represented in grey) was designed to amplify an 80-bp fragment of FXN within intron 1 and exon 2. Human mammary epithelial cell (HMEC) with copy numbers of two served as a calibrator. Error bars=s.d. n =2. (B) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin-labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8sR cells. YG8sR showed one hybridisation signal, indicating a single integration site of the FXN transgene containing one copy of the FXN gene. Scale bars: 10 μm.

    Article Snippet: The probes were prepared using purified DNA from RP11-265B8 and RP11-876N18 BAC clones, which were labelled by nick translation with biotin and digoxigenin, respectively, according to the manufacturer’s instructions (Roche).

    Techniques: Transgenic Assay, Fluorescence In Situ Hybridization, Staining, Hybridization

    Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three hybridisation signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.

    Journal: PLoS ONE

    Article Title: Cellular, Molecular and Functional Characterisation of YAC Transgenic Mouse Models of Friedreich Ataxia

    doi: 10.1371/journal.pone.0107416

    Figure Lengend Snippet: Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three hybridisation signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.

    Article Snippet: The labeled DNAs were ethanol precipitated together with Cot-1 human DNA (Roche) and resuspended in 10 µl of hybridisation buffer (Sigma).

    Techniques: Negative Control, Transgenic Assay, Fluorescence In Situ Hybridization, Staining, Hybridization

    Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three hybridisation signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.

    Journal: PLoS ONE

    Article Title: Cellular, Molecular and Functional Characterisation of YAC Transgenic Mouse Models of Friedreich Ataxia

    doi: 10.1371/journal.pone.0107416

    Figure Lengend Snippet: Transgene copy number. ( a ) Two TaqMan copy number assays were applied; Hs05092416-cn assay, represented in black, was designed to amplify a 106 bp fragment of FXN within intron 3 and Hs02407730-cn assay, represented in grey, was designed to amplify an 80 bp fragment of FXN within intron 1 and exon 2. Wild type (WT) served as a negative control with no copy number. Error bars = SD. n = 2. ( b and c ) Determination of the integration site of the transgenic FXN gene by FISH. Biotin-labelled RP11-265B8 and digoxigenin- labelled RP11-876N18 probes were hybridised onto interphase and metaphase chromosomes (DAPI stained) of YG8R, YG22R and Y47R cells. ( b ) All three cell lines showed a single integration site of the FXN transgene by metaphase analysis. ( c ) YG8R showed three hybridisation signals corresponded to the transgenic FXN , whereas YG22R and Y47R showed one signal indicating one copy of the FXN transgene. Scale bare = 10 µm.

    Article Snippet: The RP11-876N18 was detected with mouse anti-digoxigenin antibody (Sigma-Aldrich) followed by rabbit anti- mouse-FITC and anti- rabbit-FITC (Sigma-Aldrich).

    Techniques: Negative Control, Transgenic Assay, Fluorescence In Situ Hybridization, Staining, Hybridization

    Regulatory effects of a putative Oct-1 binding site located within conserved region 1. (A) Two mutant constructs were generated by site directed mutagenesis within a putative Oct-1 binding site within the identified 17 bp nucleotide sequence. MUT 1 contains one additional extra base in the middle of the sequence and MUT 2 contains two bases substitution at the start of the sequence. (B) MUT 1 and MUT 2 were generated in the pGL3-Basic::5023 construct. Luciferase expression of the mutant constructs was analyzed following co-transfection of BE(2)-M17 cells with pGL3-Basic derivatives and Oct-1 transcription factor-encoding plasmid pCMV-AC::Oct-1. The overall levels of expression were then compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p

    Journal: PLoS ONE

    Article Title: Long Range Regulation of Human FXN Gene Expression

    doi: 10.1371/journal.pone.0022001

    Figure Lengend Snippet: Regulatory effects of a putative Oct-1 binding site located within conserved region 1. (A) Two mutant constructs were generated by site directed mutagenesis within a putative Oct-1 binding site within the identified 17 bp nucleotide sequence. MUT 1 contains one additional extra base in the middle of the sequence and MUT 2 contains two bases substitution at the start of the sequence. (B) MUT 1 and MUT 2 were generated in the pGL3-Basic::5023 construct. Luciferase expression of the mutant constructs was analyzed following co-transfection of BE(2)-M17 cells with pGL3-Basic derivatives and Oct-1 transcription factor-encoding plasmid pCMV-AC::Oct-1. The overall levels of expression were then compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p

    Article Snippet: PCR products were gel extracted, digested with Hin dIII and Bgl II and cloned into the same sites of the pGL3-Basic vector (Promega, Alexandria, NSW, Australia).

    Techniques: Binding Assay, Mutagenesis, Construct, Generated, Sequencing, Luciferase, Expressing, Cotransfection, Plasmid Preparation

    DNA fragments utilized in expression analyses. Schematic representation of PCR-amplified fragments containing the FXN promoter and conserved regions cloned into the pGL3-Basic vector. The size of each fragment (bp) and the inclusion of particular conserved regions are shown. The location of each of the identified conserved non-coding regions, their distance (kb) from exon 1 of the FXN gene and their coordinates (UCSC Genome Browser, Human March 2006 assembly) are indicated. (Not to scale.)

    Journal: PLoS ONE

    Article Title: Long Range Regulation of Human FXN Gene Expression

    doi: 10.1371/journal.pone.0022001

    Figure Lengend Snippet: DNA fragments utilized in expression analyses. Schematic representation of PCR-amplified fragments containing the FXN promoter and conserved regions cloned into the pGL3-Basic vector. The size of each fragment (bp) and the inclusion of particular conserved regions are shown. The location of each of the identified conserved non-coding regions, their distance (kb) from exon 1 of the FXN gene and their coordinates (UCSC Genome Browser, Human March 2006 assembly) are indicated. (Not to scale.)

    Article Snippet: PCR products were gel extracted, digested with Hin dIII and Bgl II and cloned into the same sites of the pGL3-Basic vector (Promega, Alexandria, NSW, Australia).

    Techniques: Expressing, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    Regulatory effects of conserved region 1 on the expression of the FXN promoter. DNA fragments containing the FXN promoter and segments of conserved region 1 were inserted into the pGL3-Basic vector and luciferase expression was assayed in HeLa and BE(2)-M17 cells. The overall levels of expression were compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. (A) Fragments terminating 5,023–5,443 bp upstream of FXN exon 1; (B) Fragments terminating 4,944–5,023 bp upstream of FXN exon 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p

    Journal: PLoS ONE

    Article Title: Long Range Regulation of Human FXN Gene Expression

    doi: 10.1371/journal.pone.0022001

    Figure Lengend Snippet: Regulatory effects of conserved region 1 on the expression of the FXN promoter. DNA fragments containing the FXN promoter and segments of conserved region 1 were inserted into the pGL3-Basic vector and luciferase expression was assayed in HeLa and BE(2)-M17 cells. The overall levels of expression were compared to the pGL3-Basic::5577 vector, the expression value of which was set as 1. (A) Fragments terminating 5,023–5,443 bp upstream of FXN exon 1; (B) Fragments terminating 4,944–5,023 bp upstream of FXN exon 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p

    Article Snippet: PCR products were gel extracted, digested with Hin dIII and Bgl II and cloned into the same sites of the pGL3-Basic vector (Promega, Alexandria, NSW, Australia).

    Techniques: Expressing, Plasmid Preparation, Luciferase

    Regulatory effects of upstream conserved regions on the expression of the FXN promoter. DNA fragments containing the FXN promoter and one or multiple upstream conserved regions were inserted into the pGL3-Basic vector. The nomenclature of the vectors is based on the size of the cloned DNA fragments (bp) comprised of the sequence upstream of exon 1 of the FXN gene. Luciferase expression was assayed in HeLa and BE(2)-M17 cells. The overall levels of expression were compared to the pGL3-Basic::1269 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p

    Journal: PLoS ONE

    Article Title: Long Range Regulation of Human FXN Gene Expression

    doi: 10.1371/journal.pone.0022001

    Figure Lengend Snippet: Regulatory effects of upstream conserved regions on the expression of the FXN promoter. DNA fragments containing the FXN promoter and one or multiple upstream conserved regions were inserted into the pGL3-Basic vector. The nomenclature of the vectors is based on the size of the cloned DNA fragments (bp) comprised of the sequence upstream of exon 1 of the FXN gene. Luciferase expression was assayed in HeLa and BE(2)-M17 cells. The overall levels of expression were compared to the pGL3-Basic::1269 vector, the expression value of which was set as 1. Assays were performed in triplicate on at least three independent occasions. Error bars represent standard error of the mean. * p

    Article Snippet: PCR products were gel extracted, digested with Hin dIII and Bgl II and cloned into the same sites of the pGL3-Basic vector (Promega, Alexandria, NSW, Australia).

    Techniques: Expressing, Plasmid Preparation, Clone Assay, Sequencing, Luciferase