strain top10  (Thermo Fisher)


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    Structured Review

    Thermo Fisher strain top10
    Strain Top10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/strain top10/product/Thermo Fisher
    Average 90 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    strain top10 - by Bioz Stars, 2020-04
    90/100 stars

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    Related Articles

    Acetylene Reduction Assay:

    Article Title: Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism *
    Article Snippet: .. The E. coli strains used in this study are derivatives of strain TOP10 (F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), φ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1 ) from Invitrogen. .. Deletion mutagenesis was performed using the λ-red recombination system ( ).

    Article Title: Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)
    Article Snippet: .. E. coli strain CWG1217 [F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), ϕ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1, Δ( wzx-glf-wbbHIJK )] used for cloning is a derivative of strain TOP10 from Invitrogen that lacks part of the K-12 wb* gene cluster. .. Protein expression was performed in E. coli BL21 (DE3) [F– ompT gal dcm hsdS B (rB − mB − ) λ(DE3)] or the methionine auxotroph E. coli B834 (DE3) [F− ompT gal dcm hsdS B (rB − mB − ) met λ(DE3)].

    Amplification:

    Article Title: Specific Silencing of L392V PSEN1 Mutant Allele by RNA Interference
    Article Snippet: Reactions were performed in the following conditions—RT: 45°C 30 min, 95°C 15 min, and PCR: amplification 40 cycles (95°C 30 s; 55°C 30 s; 68°C 2 min) and termination at 72°C for 10 min. Sequences of primers were as follow: (Fow1) 5′-AAAAAA GAATTCAGATCT ATGACAGAGTTACCTGCAC-3′ and (Rev1) 5′-AAAAAA GGATCC CTAGATATAAAATTGATGGAATGC-3′. .. The cloning process was carried out using E. coli competent cells, strain TOP10 (Invitrogen).

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: PCR products were obtained with the Expand high-fidelity PCR system and amplified with the universal U3-endstop (5′-AGATTGGGGACCACTTTGTACAAGAAAGCTGGGTCTCCACCTCCGGATC-3′) and U5 primers (5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCGAAGGAGATAGAACCATG-3′) to create the recombinant site AttB. .. The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock.

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium. .. The expression cassettes of artificial miRNAs from pcDNA 6.2-GW/miR plasmids were amplified by polymerase chain reaction (PCR) and restriction enzyme sites Mfe I on the 5′ UTR and Pst I on the 3′ UTR were added, respectively, then subcloned into the pShuttle vector (sense, 5′-GCT-GCA-ATT-GGA-TGT-ACG-GGC-CAG-ATA-TAC-G-3′; antisense, 5′- CCA-GCT-GCA-GGC-TAT-GAC-CAT-GTA-ATA-CG-3′).

    Article Title: Protein-Protein Interactions in Assembly of Lipoic Acid on the 2-Oxoacid Dehydrogenases of Aerobic Metabolism *
    Article Snippet: Strain Top10 was from Invitrogen. .. Cassettes encoding resistance to either kanamycin or chloramphenicol were amplified by PCR using plasmids pKD3 and pKD4 and electroporated into induced electrocompetent MC1061 cells transformed with pKD46.

    Article Title: Mesencephalic Astrocyte-derived Neurotrophic Factor (MANF) Secretion and Cell Surface Binding Are Modulated by KDEL Receptors *
    Article Snippet: The coding sequence for enhanced GFP (Clontech) was amplified using attB-linked oligos and cloned into the EagI site of the pDONR221-MANF entry vector to produce pDONR221-GFP-MANF using Infusion (Clontech). .. All entry vectors were sequence-verified and maintained in the bacterial strain top10 (Invitrogen) prior to recombination into the pLenti6.3-V5/DEST destination vector using a standard LR reaction (Invitrogen).

    Stable Transfection:

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: Paragraph title: Plasmid construction and stable transfection ... The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada).

    Clone Assay:

    Article Title: Specific Silencing of L392V PSEN1 Mutant Allele by RNA Interference
    Article Snippet: .. The cloning process was carried out using E. coli competent cells, strain TOP10 (Invitrogen). .. Site-directed mutagenesis for introducing a point mutation in the PSEN1 gene (C→G at the position 1174) was performed using QuikChange Site-Directed Mutagenesis Kit (Stratagene) in the conditions recommended by the manufacturer.

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: Amplification was performed using the Expand high-fidelity PCR system (Roche), and the amplified fragment was cloned in front of the GFP and GUS coding sequence in the pBI101 derived gateway vector ( ). .. The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock.

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: .. The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada). .. The recombinant plasmid was purified from positive colonies by MaxiPrep, followed by phenol-chloroform extraction and ethanol precipitation.

    Article Title: Molecular Epidemiologic Identification of Escherichia coli Genes That Are Potentially Involved in Movement of the Organism from the Intestinal Tract to the Vagina and Bladder
    Article Snippet: .. Strain TOP10 (Invitrogen, San Diego, CA) was used as the host strain for recombinant clones. .. Purification, rare-cutter restriction, and pulsed-field gel electrophoresis (PFGE) of minimally sheared E. coli DNAs were performed as previously described ( ).

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Nine appropriate sequences of single-stranded oligonucleotides were generated and annealed, and respectively cloned into the miRNA expression vector pcDNA 6.2-GW/miR (Invitrogen, Carlsbad, CA) following the protocol of BLOCK-iT Pol II miR RNAi Expression Vector Kit (Invitrogen). .. Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium.

    Article Title: Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)
    Article Snippet: .. E. coli strain CWG1217 [F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), ϕ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1, Δ( wzx-glf-wbbHIJK )] used for cloning is a derivative of strain TOP10 from Invitrogen that lacks part of the K-12 wb* gene cluster. .. Protein expression was performed in E. coli BL21 (DE3) [F– ompT gal dcm hsdS B (rB − mB − ) λ(DE3)] or the methionine auxotroph E. coli B834 (DE3) [F− ompT gal dcm hsdS B (rB − mB − ) met λ(DE3)].

    Article Title: Mesencephalic Astrocyte-derived Neurotrophic Factor (MANF) Secretion and Cell Surface Binding Are Modulated by KDEL Receptors *
    Article Snippet: The coding sequence for enhanced GFP (Clontech) was amplified using attB-linked oligos and cloned into the EagI site of the pDONR221-MANF entry vector to produce pDONR221-GFP-MANF using Infusion (Clontech). .. All entry vectors were sequence-verified and maintained in the bacterial strain top10 (Invitrogen) prior to recombination into the pLenti6.3-V5/DEST destination vector using a standard LR reaction (Invitrogen).

    Construct:

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock. .. The vectors pMDC43/ATNRT2.7 were generated with GFP6 in N-terminal fusion, and the binary vectors, containing the Pro35S:GFP-ATNRT2.7 construct, were sequenced before transformation of A. tumefaciens .

    Article Title: Protein-Protein Interactions in Assembly of Lipoic Acid on the 2-Oxoacid Dehydrogenases of Aerobic Metabolism *
    Article Snippet: Strain Top10 was from Invitrogen. .. Strain MC1061 was from our laboratory collection and was used to construct gene deletions using the λ red recombinase method ( ).

    Expressing:

    Article Title: Convergent Evolution of a New Arsenic Binding Site in the ArsR/SmtB Family of Metalloregulators
    Article Snippet: .. Strain Top10 (Invitrogen) was used for protein expression. .. For expression of AfArsR from A. ferrooxidans in E. coli , plasmid pBAD arsR , in which the arsR gene is under the control of the arabinose promoter and has the sequence for a C-terminal His tag, was constructed.

    Article Title: Effects of Environmental Changes on Expression of the Oligopeptide Permease (opp) Genes of Borrelia burgdorferi
    Article Snippet: .. E. coli bacteria of strain Top10 (Invitrogen) were employed as host cells for the expression of β-galactosidase genes from reporter plasmids. .. Five oppA upstream regions were individually cloned from B. burgdorferi B31 DNA through PCR using specific primers with Bam HI and Bgl II restriction sites.

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Nine appropriate sequences of single-stranded oligonucleotides were generated and annealed, and respectively cloned into the miRNA expression vector pcDNA 6.2-GW/miR (Invitrogen, Carlsbad, CA) following the protocol of BLOCK-iT Pol II miR RNAi Expression Vector Kit (Invitrogen). .. Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium.

    Article Title: Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)
    Article Snippet: E. coli strain CWG1217 [F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), ϕ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1, Δ( wzx-glf-wbbHIJK )] used for cloning is a derivative of strain TOP10 from Invitrogen that lacks part of the K-12 wb* gene cluster. .. Protein expression was performed in E. coli BL21 (DE3) [F– ompT gal dcm hsdS B (rB − mB − ) λ(DE3)] or the methionine auxotroph E. coli B834 (DE3) [F− ompT gal dcm hsdS B (rB − mB − ) met λ(DE3)].

    Article Title: Following the Path of a Twin-arginine Precursor along the TatABC Translocase of Escherichia coli *
    Article Snippet: Strain M15pRep4 (Quiagen) transformed with pQE60-TorD ( ) was used for the expression of TorD. .. Plasmid pDULE- p Bpa ( ) was transformed into strain Top10 (Invitrogen).

    Modification:

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Human embryonic kidney 293 (HEK 293) cells, human hepatocellular carcinoma cells (Huh7), and human cervical cancer cells (Hela) were grown in Dulbecco's modified Eagle's medium medium with 10% fetal bovine serum and 100 units/ml of penicillin. .. Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium.

    Western Blot:

    Article Title: Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism *
    Article Snippet: The E. coli strains used in this study are derivatives of strain TOP10 (F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), φ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1 ) from Invitrogen. .. To remove part of the K-12 wb * gene cluster in E. coli Top10 to create E. coli CWG1217 (Δ wzx-wbbK ), pKD4 ( ) was used as a template to amplify the kanamycin resistance cassette using oligonucleotide primers containing 50 nucleotides of homology with wzx and wbbK genes (identified by lowercase); the primer sequences were 5′-acgaataaattatctttaagaagaaacgttatatatctggctgtcgttcaGTGTAGGCTGGAGCTGCTTC-3′ and 5′-atgttcttcagtaataaaattaactagttcatcaaacccaactaatacatCATATGAATATCCTCCTTAG-3′.

    Article Title: Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)
    Article Snippet: .. E. coli strain CWG1217 [F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), ϕ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1, Δ( wzx-glf-wbbHIJK )] used for cloning is a derivative of strain TOP10 from Invitrogen that lacks part of the K-12 wb* gene cluster. .. Protein expression was performed in E. coli BL21 (DE3) [F– ompT gal dcm hsdS B (rB − mB − ) λ(DE3)] or the methionine auxotroph E. coli B834 (DE3) [F− ompT gal dcm hsdS B (rB − mB − ) met λ(DE3)].

    Transformation Assay:

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock. .. The vectors pMDC43/ATNRT2.7 were generated with GFP6 in N-terminal fusion, and the binary vectors, containing the Pro35S:GFP-ATNRT2.7 construct, were sequenced before transformation of A. tumefaciens .

    Article Title: Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism *
    Article Snippet: The E. coli strains used in this study are derivatives of strain TOP10 (F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), φ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1 ) from Invitrogen. .. Electrocompetent cells of E. coli Top10 containing pSIM6 ( ) were transformed with the linear PCR product, and after recovery overnight with aeration at 30 °C, mutants were selected on LB agar containing 15 μg ml−1 kanamycin grown for 2 days at 30 °C.

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: .. Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium. .. Construction and production of recombinant adenoviruses.

    Article Title: Protein-Protein Interactions in Assembly of Lipoic Acid on the 2-Oxoacid Dehydrogenases of Aerobic Metabolism *
    Article Snippet: Strain Top10 was from Invitrogen. .. Cassettes encoding resistance to either kanamycin or chloramphenicol were amplified by PCR using plasmids pKD3 and pKD4 and electroporated into induced electrocompetent MC1061 cells transformed with pKD46.

    Article Title: Following the Path of a Twin-arginine Precursor along the TatABC Translocase of Escherichia coli *
    Article Snippet: .. Plasmid pDULE- p Bpa ( ) was transformed into strain Top10 (Invitrogen). .. Plasmid Construction —PCR-based site-specific mutations were introduced using the QuikChange Site-directed Mutagenesis kit system (Stratagene) following the manufacturer's instruction.

    Derivative Assay:

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: Amplification was performed using the Expand high-fidelity PCR system (Roche), and the amplified fragment was cloned in front of the GFP and GUS coding sequence in the pBI101 derived gateway vector ( ). .. The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock.

    Hybridization:

    Article Title: Molecular Epidemiologic Identification of Escherichia coli Genes That Are Potentially Involved in Movement of the Organism from the Intestinal Tract to the Vagina and Bladder
    Article Snippet: Three completely sequenced strains, uropathogenic E. coli CFT073 , hemorrhagic E. coli O157:H7 EDL933 , and the lab E. coli K-12 strain MG1655 , were used as controls for the hybridization. .. Strain TOP10 (Invitrogen, San Diego, CA) was used as the host strain for recombinant clones.

    Transfection:

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada). .. A total of 1 × 106 MCF7 and HEK-293 cells were then transfected in six-well plates with the recombinant plasmid (1 and 4 μg, respectively) by using Lipofectamine 2000 (Invitrogen).

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: .. Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium. .. Construction and production of recombinant adenoviruses.

    Sequencing:

    Article Title: Specific Silencing of L392V PSEN1 Mutant Allele by RNA Interference
    Article Snippet: Construction of Plasmids Coding EGFP-PSEN1 Fusion Gene Human Presenilin 1 (PSEN1 , GenBank NM_000021) coding sequence containing 1404 base pair (bp) was isolated from a total RNA, extracted from human cells (SH-SY5Y, human neuroblastoma) by reverse transcription (RT) and polymerase chain reaction (PCR) using One Step RT-PCR Kit (Qiagen). .. The cloning process was carried out using E. coli competent cells, strain TOP10 (Invitrogen).

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: Amplification was performed using the Expand high-fidelity PCR system (Roche), and the amplified fragment was cloned in front of the GFP and GUS coding sequence in the pBI101 derived gateway vector ( ). .. The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock.

    Article Title: Mesencephalic Astrocyte-derived Neurotrophic Factor (MANF) Secretion and Cell Surface Binding Are Modulated by KDEL Receptors *
    Article Snippet: .. All entry vectors were sequence-verified and maintained in the bacterial strain top10 (Invitrogen) prior to recombination into the pLenti6.3-V5/DEST destination vector using a standard LR reaction (Invitrogen). .. All plasmids based on LV backbones were maintained in the recombination-deficient bacterial stain Stbl3 (Invitrogen).

    Synthesized:

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: Specific primers for the 17β-HSD1 gene (gene HSD17B1; GenBank accession no. ) were synthesized and used in PCR to amplify human 17β-HSD1 cDNA that had been previously subcloned ( , ). .. The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada).

    Introduce:

    Article Title: Specific Silencing of L392V PSEN1 Mutant Allele by RNA Interference
    Article Snippet: Primers were designed to introduce EcoR I, Bgl II restriction sites to the 5′ end and BamH I restriction site to the 3′ end of PSEN1 gene, appropriate for cloning into pUC18 (Invitrogen) and pEGFP-C1 (BD Biosciences) plasmids. .. The cloning process was carried out using E. coli competent cells, strain TOP10 (Invitrogen).

    Generated:

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock. .. The vectors pMDC43/ATNRT2.7 were generated with GFP6 in N-terminal fusion, and the binary vectors, containing the Pro35S:GFP-ATNRT2.7 construct, were sequenced before transformation of A. tumefaciens .

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Nine appropriate sequences of single-stranded oligonucleotides were generated and annealed, and respectively cloned into the miRNA expression vector pcDNA 6.2-GW/miR (Invitrogen, Carlsbad, CA) following the protocol of BLOCK-iT Pol II miR RNAi Expression Vector Kit (Invitrogen). .. Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Specific Silencing of L392V PSEN1 Mutant Allele by RNA Interference
    Article Snippet: Construction of Plasmids Coding EGFP-PSEN1 Fusion Gene Human Presenilin 1 (PSEN1 , GenBank NM_000021) coding sequence containing 1404 base pair (bp) was isolated from a total RNA, extracted from human cells (SH-SY5Y, human neuroblastoma) by reverse transcription (RT) and polymerase chain reaction (PCR) using One Step RT-PCR Kit (Qiagen). .. The cloning process was carried out using E. coli competent cells, strain TOP10 (Invitrogen).

    Recombinant:

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: PCR products were obtained with the Expand high-fidelity PCR system and amplified with the universal U3-endstop (5′-AGATTGGGGACCACTTTGTACAAGAAAGCTGGGTCTCCACCTCCGGATC-3′) and U5 primers (5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCGAAGGAGATAGAACCATG-3′) to create the recombinant site AttB. .. The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock.

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada). .. The recombinant plasmid was purified from positive colonies by MaxiPrep, followed by phenol-chloroform extraction and ethanol precipitation.

    Article Title: Molecular Epidemiologic Identification of Escherichia coli Genes That Are Potentially Involved in Movement of the Organism from the Intestinal Tract to the Vagina and Bladder
    Article Snippet: .. Strain TOP10 (Invitrogen, San Diego, CA) was used as the host strain for recombinant clones. .. Purification, rare-cutter restriction, and pulsed-field gel electrophoresis (PFGE) of minimally sheared E. coli DNAs were performed as previously described ( ).

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium. .. Construction and production of recombinant adenoviruses.

    Cellular Antioxidant Activity Assay:

    Article Title: Following the Path of a Twin-arginine Precursor along the TatABC Translocase of Escherichia coli *
    Article Snippet: Plasmid pDULE- p Bpa ( ) was transformed into strain Top10 (Invitrogen). .. For position 15 (Phe) the two complementary oligonucleotides TorA-PhoAF15for (5′-CA TCA CGT CGG CGT TAG CTG GCA CAA CTC GGC GGC-3′) and TorA-PhoAF15rev(5′-GCCGCCGAGTTGTGCCAGCTAACG CCG ACG TGA TG-3′) were used to give the plasmid pET28aTorA-PhoAF15.

    In Vivo:

    Article Title: Convergent Evolution of a New Arsenic Binding Site in the ArsR/SmtB Family of Metalloregulators
    Article Snippet: Plasmids pB2 lacZ , pB3 lacZ , and E. coli strain ACSH501q ( rspl Δ( lac-pro ) (F′ tra D36 pro AB lacIq ΔM15) Δ ars::cam ) ( ) were used for analysis of AfArsR function in vivo . .. Strain Top10 (Invitrogen) was used for protein expression.

    Mutagenesis:

    Article Title: Specific Silencing of L392V PSEN1 Mutant Allele by RNA Interference
    Article Snippet: The cloning process was carried out using E. coli competent cells, strain TOP10 (Invitrogen). .. Site-directed mutagenesis for introducing a point mutation in the PSEN1 gene (C→G at the position 1174) was performed using QuikChange Site-Directed Mutagenesis Kit (Stratagene) in the conditions recommended by the manufacturer.

    Article Title: Convergent Evolution of a New Arsenic Binding Site in the ArsR/SmtB Family of Metalloregulators
    Article Snippet: E. coli strains DH5 α (Promega, Madison, WI), XL1-Blue (F′ proA + B + lacIq Δ( lacZ ) M15 Tn 10 ( tetR ) recA1 endA1 gyr96 thi-1 hsdR17 supE44 relA1 lac ), were used for plasmid construction, replication, and mutagenesis. .. Strain Top10 (Invitrogen) was used for protein expression.

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock. .. Arabidopsis plants, the wild type or the atnrt2.1-1 mutant, were transformed according to the in planta method using the surfactant Silwet L-77 ( ).

    Article Title: Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism *
    Article Snippet: The E. coli strains used in this study are derivatives of strain TOP10 (F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), φ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1 ) from Invitrogen. .. Deletion mutagenesis was performed using the λ-red recombination system ( ).

    Article Title: Protein-Protein Interactions in Assembly of Lipoic Acid on the 2-Oxoacid Dehydrogenases of Aerobic Metabolism *
    Article Snippet: Strain Top10 was from Invitrogen. .. Using this method, Δ aceE , Δ aceF , Δ aceEF , and Δ lpd mutant strains carrying the chloramphenicol resistance cassette in place of the coding sequences were obtained.

    Article Title: Following the Path of a Twin-arginine Precursor along the TatABC Translocase of Escherichia coli *
    Article Snippet: Plasmid pDULE- p Bpa ( ) was transformed into strain Top10 (Invitrogen). .. Plasmid Construction —PCR-based site-specific mutations were introduced using the QuikChange Site-directed Mutagenesis kit system (Stratagene) following the manufacturer's instruction.

    Isolation:

    Article Title: Specific Silencing of L392V PSEN1 Mutant Allele by RNA Interference
    Article Snippet: Construction of Plasmids Coding EGFP-PSEN1 Fusion Gene Human Presenilin 1 (PSEN1 , GenBank NM_000021) coding sequence containing 1404 base pair (bp) was isolated from a total RNA, extracted from human cells (SH-SY5Y, human neuroblastoma) by reverse transcription (RT) and polymerase chain reaction (PCR) using One Step RT-PCR Kit (Qiagen). .. The cloning process was carried out using E. coli competent cells, strain TOP10 (Invitrogen).

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: First primers AttB1-ATNRT2.7 start (AttB1, 5′-GAGCCATCTCAACGCAAC-3′) and AttB2-ATNRT2.7 End-Stop (AttB2, 5′-AACAAACGGGACGTAGACTACC-3′) were used to amplify a complete ATNRT2.7 cDNA from our previously isolated clone (see above). .. The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock.

    Purification:

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada). .. The recombinant plasmid was purified from positive colonies by MaxiPrep, followed by phenol-chloroform extraction and ethanol precipitation.

    Polymerase Chain Reaction:

    Article Title: Specific Silencing of L392V PSEN1 Mutant Allele by RNA Interference
    Article Snippet: Reactions were performed in the following conditions—RT: 45°C 30 min, 95°C 15 min, and PCR: amplification 40 cycles (95°C 30 s; 55°C 30 s; 68°C 2 min) and termination at 72°C for 10 min. Sequences of primers were as follow: (Fow1) 5′-AAAAAA GAATTCAGATCT ATGACAGAGTTACCTGCAC-3′ and (Rev1) 5′-AAAAAA GGATCC CTAGATATAAAATTGATGGAATGC-3′. .. The cloning process was carried out using E. coli competent cells, strain TOP10 (Invitrogen).

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: PCR products were obtained with the Expand high-fidelity PCR system and amplified with the universal U3-endstop (5′-AGATTGGGGACCACTTTGTACAAGAAAGCTGGGTCTCCACCTCCGGATC-3′) and U5 primers (5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCGAAGGAGATAGAACCATG-3′) to create the recombinant site AttB. .. The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock.

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: Specific primers for the 17β-HSD1 gene (gene HSD17B1; GenBank accession no. ) were synthesized and used in PCR to amplify human 17β-HSD1 cDNA that had been previously subcloned ( , ). .. The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada).

    Article Title: Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism *
    Article Snippet: The E. coli strains used in this study are derivatives of strain TOP10 (F− , mcrA , Δ( mrr-hsdRMS-mcrBC ), φ80, lacZ Δ M15 , Δ lacX74 , deoR , nupG , recA1 , araD139 , Δ( ara-leu )7697, galU , galK , rpsL (Strr ), endA1 ) from Invitrogen. .. Electrocompetent cells of E. coli Top10 containing pSIM6 ( ) were transformed with the linear PCR product, and after recovery overnight with aeration at 30 °C, mutants were selected on LB agar containing 15 μg ml−1 kanamycin grown for 2 days at 30 °C.

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium. .. The expression cassettes of artificial miRNAs from pcDNA 6.2-GW/miR plasmids were amplified by polymerase chain reaction (PCR) and restriction enzyme sites Mfe I on the 5′ UTR and Pst I on the 3′ UTR were added, respectively, then subcloned into the pShuttle vector (sense, 5′-GCT-GCA-ATT-GGA-TGT-ACG-GGC-CAG-ATA-TAC-G-3′; antisense, 5′- CCA-GCT-GCA-GGC-TAT-GAC-CAT-GTA-ATA-CG-3′).

    Article Title: Protein-Protein Interactions in Assembly of Lipoic Acid on the 2-Oxoacid Dehydrogenases of Aerobic Metabolism *
    Article Snippet: Strain Top10 was from Invitrogen. .. Cassettes encoding resistance to either kanamycin or chloramphenicol were amplified by PCR using plasmids pKD3 and pKD4 and electroporated into induced electrocompetent MC1061 cells transformed with pKD46.

    Blocking Assay:

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Nine appropriate sequences of single-stranded oligonucleotides were generated and annealed, and respectively cloned into the miRNA expression vector pcDNA 6.2-GW/miR (Invitrogen, Carlsbad, CA) following the protocol of BLOCK-iT Pol II miR RNAi Expression Vector Kit (Invitrogen). .. Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium.

    Plasmid Preparation:

    Article Title: Convergent Evolution of a New Arsenic Binding Site in the ArsR/SmtB Family of Metalloregulators
    Article Snippet: E. coli strains DH5 α (Promega, Madison, WI), XL1-Blue (F′ proA + B + lacIq Δ( lacZ ) M15 Tn 10 ( tetR ) recA1 endA1 gyr96 thi-1 hsdR17 supE44 relA1 lac ), were used for plasmid construction, replication, and mutagenesis. .. Strain Top10 (Invitrogen) was used for protein expression.

    Article Title: The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds [W]
    Article Snippet: Amplification was performed using the Expand high-fidelity PCR system (Roche), and the amplified fragment was cloned in front of the GFP and GUS coding sequence in the pBI101 derived gateway vector ( ). .. The product of recombination reactions (BP reactions) was used to transform competent Escherichia coli , strain TOP10 (Invitrogen), by heat shock.

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: .. The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada). .. The recombinant plasmid was purified from positive colonies by MaxiPrep, followed by phenol-chloroform extraction and ethanol precipitation.

    Article Title: Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Article Snippet: Nine appropriate sequences of single-stranded oligonucleotides were generated and annealed, and respectively cloned into the miRNA expression vector pcDNA 6.2-GW/miR (Invitrogen, Carlsbad, CA) following the protocol of BLOCK-iT Pol II miR RNAi Expression Vector Kit (Invitrogen). .. Transfections were carried out using Lipofectamine 2000 (Invitrogen) for HEK 293 cells or Lipofectamine LTX (Invitrogen) for Huh7 cells and Hela cells, according to the manufacturer's instructions. pcDNA 6.2-GW/miR plasmids and pShuttle plasmids (Clonetech, Mountain View, CA) were transformed into the bacterial strain TOP10 (Invitrogen) at 37 °C in S.O.C medium.

    Article Title: Following the Path of a Twin-arginine Precursor along the TatABC Translocase of Escherichia coli *
    Article Snippet: .. Plasmid pDULE- p Bpa ( ) was transformed into strain Top10 (Invitrogen). .. Plasmid Construction —PCR-based site-specific mutations were introduced using the QuikChange Site-directed Mutagenesis kit system (Stratagene) following the manufacturer's instruction.

    Article Title: Mesencephalic Astrocyte-derived Neurotrophic Factor (MANF) Secretion and Cell Surface Binding Are Modulated by KDEL Receptors *
    Article Snippet: .. All entry vectors were sequence-verified and maintained in the bacterial strain top10 (Invitrogen) prior to recombination into the pLenti6.3-V5/DEST destination vector using a standard LR reaction (Invitrogen). .. All plasmids based on LV backbones were maintained in the recombination-deficient bacterial stain Stbl3 (Invitrogen).

    Ethanol Precipitation:

    Article Title: 17β-Hydroxysteroid Dehydrogenase Type 1 Stimulates Breast Cancer by Dihydrotestosterone Inactivation in Addition to Estradiol Production
    Article Snippet: The fragment thus obtained was inserted in the Hin dIII/ Eco RI site of the pcDNA3.1(+) vector and cloned in bacterial strain TOP10 (Invitrogen, Burlington, Ontario, Canada). .. The recombinant plasmid was purified from positive colonies by MaxiPrep, followed by phenol-chloroform extraction and ethanol precipitation.

    Produced:

    Article Title: Mesencephalic Astrocyte-derived Neurotrophic Factor (MANF) Secretion and Cell Surface Binding Are Modulated by KDEL Receptors *
    Article Snippet: The pDONR221-SP-GFP-RTDL entry vector was produced by amplifying the SP-GFP open reading frame (from pLenti6.3-SP-GFP) using attB-linked oligos, with the reverse oligo including the nucleotides encoding the ASARTDL sequence. .. All entry vectors were sequence-verified and maintained in the bacterial strain top10 (Invitrogen) prior to recombination into the pLenti6.3-V5/DEST destination vector using a standard LR reaction (Invitrogen).

    CTG Assay:

    Article Title: Following the Path of a Twin-arginine Precursor along the TatABC Translocase of Escherichia coli *
    Article Snippet: Plasmid pDULE- p Bpa ( ) was transformed into strain Top10 (Invitrogen). .. For position 15 (Phe) the two complementary oligonucleotides TorA-PhoAF15for (5′-CA TCA CGT CGG CGT TAG CTG GCA CAA CTC GGC GGC-3′) and TorA-PhoAF15rev(5′-GCCGCCGAGTTGTGCCAGCTAACG CCG ACG TGA TG-3′) were used to give the plasmid pET28aTorA-PhoAF15.

    Staining:

    Article Title: Mesencephalic Astrocyte-derived Neurotrophic Factor (MANF) Secretion and Cell Surface Binding Are Modulated by KDEL Receptors *
    Article Snippet: All entry vectors were sequence-verified and maintained in the bacterial strain top10 (Invitrogen) prior to recombination into the pLenti6.3-V5/DEST destination vector using a standard LR reaction (Invitrogen). .. All plasmids based on LV backbones were maintained in the recombination-deficient bacterial stain Stbl3 (Invitrogen).

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