oneshot top10 competent cells  (Thermo Fisher)


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    Structured Review

    Thermo Fisher oneshot top10 competent cells
    Oneshot Top10 Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oneshot top10 competent cells/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    oneshot top10 competent cells - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Transduction:

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: To filter induced PCR mutations and distinguish possible allelic variants, at least four independent PCR products, and clones from each L1 transduction family member were capillary sequenced using 12 overlapping primer pairs ( ) distributed at ∼500-bp intervals covering the entire L1 sequence. .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    Clone Assay:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression. .. Positive clones were cultured in Luria broth (LB) media with 100 μg mL–1 ampicillin (Amp) at 37 °C and 220 rpm, plasmids were isolated and promoter integrity was confirmed by DNA sequencing.

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: .. The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA). .. Plasmids were isolated from E. coli using the QIAprep Spin MiniPrep Kit (Qiagen, Hilden Germany).

    Article Title: AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach
    Article Snippet: .. E . coli strains The following commercially available strains of E . coli were also used for AQUA Cloning: One Shot TOP10 (Invitrogen cat. No. C4040-03, competency: > 109 CFU/μg), NEB5α (NEB, cat. No. C2987I, competency: 1–3 x 109 CFU/μg), NEB10β (NEB, cat. No. C3019I, competency: 1–3 x 109 CFU/μg), BL21 (DE3) (NEB, cat. No. C2527I, competency: 1–5 x 107 CFU/μg), JM109 (Promega, cat. No. L2005, competency: 108 CFU/μg). .. Confocal imaging For confocal imaging, human embryonic kidney cells (HEK-293T) [ ], or NIH/3T3 fibroblasts cells (ATCC CRL-1658) were maintained in Dulbecco’s modified Eagle’s medium (PAN, cat. No. P03-0710) supplemented with 10% fetal bovine serum (PAN, cat. no. 1502-P123002, batch no. P123002), 100 U/mL penicillin and 0.1 mg/mL streptomycin (PAN).

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: Paragraph title: Sequencing and cloning of DGGE bands ... The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing.

    Article Title: α-Xylosidase plays essential roles in xyloglucan remodelling, maintenance of cell wall integrity, and seed germination in Arabidopsis thaliana
    Article Snippet: .. The product was cloned into the Gateway entry vector using the pENTR Directional TOPO Cloning Kit (Invitrogen, K2400-20SP) and transformed One Shot Chemically Competent Escherichia coli (Invitrogen, C4040-03). .. Kanamycin-resistant colonies were selected and plasmid DNA was prepared using the QIAprep Spin Miniprep Kit (Qiagen, 27104).

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: Paragraph title: L1 genotyping and cloning. ... Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    Article Title: Fecal Components Modulate Human Astrovirus Infectivity in Cells and Reconstituted Intestinal Tissues
    Article Snippet: .. The PCR product was cloned into a pCR4-TOPO cloning vector (Invitrogen, USA), and the plasmid was transformed into chemically competent E. coli One Shot TOP10 cells (Invitrogen, USA). .. The plasmid was in vitro transcribed using a T7 RiboMAX large-scale RNA production system (Promega), and 10-fold dilution series (from 107 to 104 copies/ml) were used as a standard.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: The TfRC1 PCR product was directionally cloned into pcDNA 3.1 vector as per manufacturer's instructions (Life Technologies). .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin.

    Amplification:

    Article Title: Salt-responsive gut commensal modulates TH17 axis and disease
    Article Snippet: .. Standard curves for quantification consisted in ten-fold serial dilutions in the range of 108 to 100 copies of E. coli (Invitrogen, C404010) or L. murinus isolate 16S rRNA gene amplified with primers 27F (5’-GTTTGATCCTGGCTCAG-3’) and 1492R (5’-CGGCTA CCTTGTTACGAC-3’) . ..

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: Products of the nested PCR were used as the template for final amplification of the synthetic promoters using external primers that corresponded to the terminal 20 bp of each sequence. .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression.

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: The resulting amplicon spanned the 3′ end of cas1 through the truncated IS1194 fragment with a repeat-s4-repeat CRISPR sandwiched in-between. .. The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA).

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: It was amplified by Phusion HF DNA polymerase (M0530L) and purified by the QIAquick PCR Purification Kit (#28106) and QIA gel Extraction kit (#28706). .. Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher).

    Article Title: α-Xylosidase plays essential roles in xyloglucan remodelling, maintenance of cell wall integrity, and seed germination in Arabidopsis thaliana
    Article Snippet: Transgenic plants The TRG1/XYL1 promoter region and the gene containing upstream and downstream regions were amplified from Ws genomic DNA by high-fidelity PCR with specific primers ( Supplementary Table S2 ) and PrimeSTAR DNA polymerase (Takara Bio Inc.). .. The product was cloned into the Gateway entry vector using the pENTR Directional TOPO Cloning Kit (Invitrogen, K2400-20SP) and transformed One Shot Chemically Competent Escherichia coli (Invitrogen, C4040-03).

    Article Title: Exome Sequencing Identifies a Novel CEACAM16 Mutation Associated with Autosomal Dominant Nonsyndromic Hearing Loss DFNA4B in a Chinese Family
    Article Snippet: .. Plasmid transfection, amplification and extraction The two kinds of plasmid pRK5-wild type (WT) and G169R mutant (MT) CEACAM16 -Flag were transformed into One Shot TOP10 competent E. coil (Life technologies) on LB culture plates with ampicillin. ..

    Article Title: Fecal Components Modulate Human Astrovirus Infectivity in Cells and Reconstituted Intestinal Tissues
    Article Snippet: For preparing a quantitative reference standard, a fragment covering the target was amplified from HAstV-1. .. The PCR product was cloned into a pCR4-TOPO cloning vector (Invitrogen, USA), and the plasmid was transformed into chemically competent E. coli One Shot TOP10 cells (Invitrogen, USA).

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: Each 800 bp region was amplified via sequencing PCR, using the BigDye Terminator mix (Life Technologies) as per manufacturer's instructions. .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin.

    DNA Ligation:

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations. .. Transformed cells were plated onto LB medium plates (100 μl plate−1 ) containing ampicillin (0.06 mg ml−1 ), isopropyl-β- d -thiogalactopyranoside (IPTG; 0.01 mg ml−1 ), and 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (0.04 mg ml−1 ) and incubated overnight at 37°C to verify DNA ligation using blue-white selection.

    TA Cloning:

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: Cloning was performed using the TA cloning vector pJET1.2 (CloneJet PCR cloning kit; Fermentas, Glen Burnie, MD, USA). .. The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing.

    Construct:

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: .. The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA). .. Plasmids were isolated from E. coli using the QIAprep Spin MiniPrep Kit (Qiagen, Hilden Germany).

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: .. The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing. .. Phylogeny of DGGE bands A phylogenetic tree was inferred from the pqqC sequences (501 bp) of (i) the DGGE bands, (ii) a selection of 14 reference pseudomonas including 11 Pseudomonas strains listed in and three GenBank sequences (P. fluorescens SBW25, GenBank database accession number NC_012660.1; P. fluorescens Pf0-1, NC_007492.2; and P. syringae pv. syringae B728a, CP000075.1), (iii) 30 RW09-C isolates , and (iv) 76 RW09-NC clones obtained from wheat roots as described in Meyer et al. (Genbank database accession numbers JN397402 - 477).

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: For each L1, a consensus sequence was obtained, and a mutation-free construct was reconstructed by performing multiple restriction enzyme digestions. .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    SYBR Green Assay:

    Article Title: Salt-responsive gut commensal modulates TH17 axis and disease
    Article Snippet: Samples were quantified in 10 μl reactions containing 1x SYBR Green Master Mix (AB), 300 nM of each primer and 4 ng of genomic DNA. .. Standard curves for quantification consisted in ten-fold serial dilutions in the range of 108 to 100 copies of E. coli (Invitrogen, C404010) or L. murinus isolate 16S rRNA gene amplified with primers 27F (5’-GTTTGATCCTGGCTCAG-3’) and 1492R (5’-CGGCTA CCTTGTTACGAC-3’) .

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: PCR product size was determined by electrophoresis at 50 V using a 0.7% agarose gel and visualized using SYBR Green Fluorescent dye. .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin.

    Incubation:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: A total of 50 pmol of forward and reverse primers with 5 units of Taq DNA polymerase were combined and incubated at 72 °C for 65 min, hybridizing the two oligos. .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression.

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations. .. Transformed cells were plated onto LB medium plates (100 μl plate−1 ) containing ampicillin (0.06 mg ml−1 ), isopropyl-β- d -thiogalactopyranoside (IPTG; 0.01 mg ml−1 ), and 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (0.04 mg ml−1 ) and incubated overnight at 37°C to verify DNA ligation using blue-white selection.

    Expressing:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression. .. Positive clones were cultured in Luria broth (LB) media with 100 μg mL–1 ampicillin (Amp) at 37 °C and 220 rpm, plasmids were isolated and promoter integrity was confirmed by DNA sequencing.

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: The PCR product was cut with NEB CutSmart and ligated overnight at 16ºC to p3XFLAG-CMV™ -7.1 expression vector (E7533 SIGMA). .. Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher).

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin. .. The insertion of the TfRC1 gene into the plasmid was verified by performing PCR on the isolated plasmid using the forward and reverse primers (T7 forward and BGH reverse) provided with the pcDNA 3.1 Directional TOPO expression kit (Life Technologies).

    Modification:

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Total genomic DNA (gDNA) of SA-01 was isolated using a modified SDS-proteinase K treatment method ( ) and restriction digested using the endonuclease Sau3AI (New England BioLabs, Beverly, MA), using serial dilutions of enzyme ranging from 250 U ml−1 to 4.6 U ml−1 (15 min at 37°C). .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations.

    Western Blot:

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher). .. The cell was post-transfected for 48 hours at 37°C, then the transfection efficacy was determined by qPCR quantification and western blot analysis.

    Transformation Assay:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression. .. Positive clones were cultured in Luria broth (LB) media with 100 μg mL–1 ampicillin (Amp) at 37 °C and 220 rpm, plasmids were isolated and promoter integrity was confirmed by DNA sequencing.

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: .. The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA). .. Plasmids were isolated from E. coli using the QIAprep Spin MiniPrep Kit (Qiagen, Hilden Germany).

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: .. Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher). .. The overexpression gene transient transfection was carried out with Lipofectamine 3000 reagent (Invitrogen, CA, USA), while transient knockdown using Mission® esiRNA (EHU018231 SIGMA) with Lipofectamine 2000 following the manufacturer's protocol.

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: .. The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing. .. Phylogeny of DGGE bands A phylogenetic tree was inferred from the pqqC sequences (501 bp) of (i) the DGGE bands, (ii) a selection of 14 reference pseudomonas including 11 Pseudomonas strains listed in and three GenBank sequences (P. fluorescens SBW25, GenBank database accession number NC_012660.1; P. fluorescens Pf0-1, NC_007492.2; and P. syringae pv. syringae B728a, CP000075.1), (iii) 30 RW09-C isolates , and (iv) 76 RW09-NC clones obtained from wheat roots as described in Meyer et al. (Genbank database accession numbers JN397402 - 477).

    Article Title: α-Xylosidase plays essential roles in xyloglucan remodelling, maintenance of cell wall integrity, and seed germination in Arabidopsis thaliana
    Article Snippet: .. The product was cloned into the Gateway entry vector using the pENTR Directional TOPO Cloning Kit (Invitrogen, K2400-20SP) and transformed One Shot Chemically Competent Escherichia coli (Invitrogen, C4040-03). .. Kanamycin-resistant colonies were selected and plasmid DNA was prepared using the QIAprep Spin Miniprep Kit (Qiagen, 27104).

    Article Title: Exome Sequencing Identifies a Novel CEACAM16 Mutation Associated with Autosomal Dominant Nonsyndromic Hearing Loss DFNA4B in a Chinese Family
    Article Snippet: .. Plasmid transfection, amplification and extraction The two kinds of plasmid pRK5-wild type (WT) and G169R mutant (MT) CEACAM16 -Flag were transformed into One Shot TOP10 competent E. coil (Life technologies) on LB culture plates with ampicillin. ..

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations. .. Transformed cells were plated onto LB medium plates (100 μl plate−1 ) containing ampicillin (0.06 mg ml−1 ), isopropyl-β- d -thiogalactopyranoside (IPTG; 0.01 mg ml−1 ), and 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (0.04 mg ml−1 ) and incubated overnight at 37°C to verify DNA ligation using blue-white selection.

    Article Title: Fecal Components Modulate Human Astrovirus Infectivity in Cells and Reconstituted Intestinal Tissues
    Article Snippet: .. The PCR product was cloned into a pCR4-TOPO cloning vector (Invitrogen, USA), and the plasmid was transformed into chemically competent E. coli One Shot TOP10 cells (Invitrogen, USA). .. The plasmid was in vitro transcribed using a T7 RiboMAX large-scale RNA production system (Promega), and 10-fold dilution series (from 107 to 104 copies/ml) were used as a standard.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin. .. Plasmids were purified from these E. coli cells using the PureLink Quick Plasmid Miniprep kit (Life Technologies) as per manufacturer's instructions.

    Over Expression:

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher). .. The overexpression gene transient transfection was carried out with Lipofectamine 3000 reagent (Invitrogen, CA, USA), while transient knockdown using Mission® esiRNA (EHU018231 SIGMA) with Lipofectamine 2000 following the manufacturer's protocol.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: Paragraph title: TfRC1 overexpression ... The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin.

    Transfection:

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: Paragraph title: Molecular cloning and gene transient transfection ... Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher).

    Article Title: Exome Sequencing Identifies a Novel CEACAM16 Mutation Associated with Autosomal Dominant Nonsyndromic Hearing Loss DFNA4B in a Chinese Family
    Article Snippet: .. Plasmid transfection, amplification and extraction The two kinds of plasmid pRK5-wild type (WT) and G169R mutant (MT) CEACAM16 -Flag were transformed into One Shot TOP10 competent E. coil (Life technologies) on LB culture plates with ampicillin. ..

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin. .. TREx Jurkat cells were transfected with the TfRC1 plasmid using Lipofectamine 2000 (Life Technologies) as per manufacturer's instructions.

    Sequencing:

    Article Title: Salt-responsive gut commensal modulates TH17 axis and disease
    Article Snippet: To optimize bacterial community structure representation, a mutanolysin (Sigma) digestion step was included (6 μl of 25kU/ml stock/sample) . qPCR on a 7500 Sequence Detector (Applied Biosystems, AB) was used to enumerate bacterial 16S rRNA gene copies in the genomic DNA extracted from stool samples. .. Standard curves for quantification consisted in ten-fold serial dilutions in the range of 108 to 100 copies of E. coli (Invitrogen, C404010) or L. murinus isolate 16S rRNA gene amplified with primers 27F (5’-GTTTGATCCTGGCTCAG-3’) and 1492R (5’-CGGCTA CCTTGTTACGAC-3’) .

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: Products of the nested PCR were used as the template for final amplification of the synthetic promoters using external primers that corresponded to the terminal 20 bp of each sequence. .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression.

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: .. The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing. .. Phylogeny of DGGE bands A phylogenetic tree was inferred from the pqqC sequences (501 bp) of (i) the DGGE bands, (ii) a selection of 14 reference pseudomonas including 11 Pseudomonas strains listed in and three GenBank sequences (P. fluorescens SBW25, GenBank database accession number NC_012660.1; P. fluorescens Pf0-1, NC_007492.2; and P. syringae pv. syringae B728a, CP000075.1), (iii) 30 RW09-C isolates , and (iv) 76 RW09-NC clones obtained from wheat roots as described in Meyer et al. (Genbank database accession numbers JN397402 - 477).

    Article Title: α-Xylosidase plays essential roles in xyloglucan remodelling, maintenance of cell wall integrity, and seed germination in Arabidopsis thaliana
    Article Snippet: The product was cloned into the Gateway entry vector using the pENTR Directional TOPO Cloning Kit (Invitrogen, K2400-20SP) and transformed One Shot Chemically Competent Escherichia coli (Invitrogen, C4040-03). .. The insertion of the entry plasmids was confirmed by sequencing with primers listed in Supplementary Table S5 .

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: For each L1, a consensus sequence was obtained, and a mutation-free construct was reconstructed by performing multiple restriction enzyme digestions. .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: Each 800 bp region was amplified via sequencing PCR, using the BigDye Terminator mix (Life Technologies) as per manufacturer's instructions. .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin.

    esiRNA:

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher). .. The overexpression gene transient transfection was carried out with Lipofectamine 3000 reagent (Invitrogen, CA, USA), while transient knockdown using Mission® esiRNA (EHU018231 SIGMA) with Lipofectamine 2000 following the manufacturer's protocol.

    Ligation:

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions. ..

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations. .. Transformed cells were plated onto LB medium plates (100 μl plate−1 ) containing ampicillin (0.06 mg ml−1 ), isopropyl-β- d -thiogalactopyranoside (IPTG; 0.01 mg ml−1 ), and 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (0.04 mg ml−1 ) and incubated overnight at 37°C to verify DNA ligation using blue-white selection.

    Transferring:

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: Briefly, the central part of DGGE bands was cut out using sterile pipette tips. .. The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing.

    Cell Culture:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression. .. Positive clones were cultured in Luria broth (LB) media with 100 μg mL–1 ampicillin (Amp) at 37 °C and 220 rpm, plasmids were isolated and promoter integrity was confirmed by DNA sequencing.

    Generated:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: An initial template was generated with two, 111 bp, overlapping synthetic oligos corresponding to the sequence of each desired promoter. .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression.

    DNA Sequencing:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression. .. Positive clones were cultured in Luria broth (LB) media with 100 μg mL–1 ampicillin (Amp) at 37 °C and 220 rpm, plasmids were isolated and promoter integrity was confirmed by DNA sequencing.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Fecal Components Modulate Human Astrovirus Infectivity in Cells and Reconstituted Intestinal Tissues
    Article Snippet: RT-qPCR was performed with a QuantiTect probe RT-PCR kit (Qiagen) in a StepOne Applied Biosystems thermocycler. .. The PCR product was cloned into a pCR4-TOPO cloning vector (Invitrogen, USA), and the plasmid was transformed into chemically competent E. coli One Shot TOP10 cells (Invitrogen, USA).

    Denaturing Gradient Gel Electrophoresis:

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: Paragraph title: Sequencing and cloning of DGGE bands ... The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing.

    DNA Extraction:

    Article Title: Salt-responsive gut commensal modulates TH17 axis and disease
    Article Snippet: Paragraph title: DNA extraction from feces and qPCR of 16S rRNA genes ... Standard curves for quantification consisted in ten-fold serial dilutions in the range of 108 to 100 copies of E. coli (Invitrogen, C404010) or L. murinus isolate 16S rRNA gene amplified with primers 27F (5’-GTTTGATCCTGGCTCAG-3’) and 1492R (5’-CGGCTA CCTTGTTACGAC-3’) .

    Molecular Cloning:

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: Paragraph title: Molecular cloning and gene transient transfection ... Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher).

    Fluorescence:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression. .. Positive clones were cultured in Luria broth (LB) media with 100 μg mL–1 ampicillin (Amp) at 37 °C and 220 rpm, plasmids were isolated and promoter integrity was confirmed by DNA sequencing.

    Mutagenesis:

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: For each L1, a consensus sequence was obtained, and a mutation-free construct was reconstructed by performing multiple restriction enzyme digestions. .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    Article Title: Exome Sequencing Identifies a Novel CEACAM16 Mutation Associated with Autosomal Dominant Nonsyndromic Hearing Loss DFNA4B in a Chinese Family
    Article Snippet: .. Plasmid transfection, amplification and extraction The two kinds of plasmid pRK5-wild type (WT) and G169R mutant (MT) CEACAM16 -Flag were transformed into One Shot TOP10 competent E. coil (Life technologies) on LB culture plates with ampicillin. ..

    Isolation:

    Article Title: Salt-responsive gut commensal modulates TH17 axis and disease
    Article Snippet: DNA extraction from feces and qPCR of 16S rRNA genes DNA from stool samples was isolated using QIAamp DNA Stool Mini Kit (Qiagen) according to the manufacturer's instructions. .. Standard curves for quantification consisted in ten-fold serial dilutions in the range of 108 to 100 copies of E. coli (Invitrogen, C404010) or L. murinus isolate 16S rRNA gene amplified with primers 27F (5’-GTTTGATCCTGGCTCAG-3’) and 1492R (5’-CGGCTA CCTTGTTACGAC-3’) .

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression. .. Positive clones were cultured in Luria broth (LB) media with 100 μg mL–1 ampicillin (Amp) at 37 °C and 220 rpm, plasmids were isolated and promoter integrity was confirmed by DNA sequencing.

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA). .. Plasmids were isolated from E. coli using the QIAprep Spin MiniPrep Kit (Qiagen, Hilden Germany).

    Article Title: Exome Sequencing Identifies a Novel CEACAM16 Mutation Associated with Autosomal Dominant Nonsyndromic Hearing Loss DFNA4B in a Chinese Family
    Article Snippet: Plasmid transfection, amplification and extraction The two kinds of plasmid pRK5-wild type (WT) and G169R mutant (MT) CEACAM16 -Flag were transformed into One Shot TOP10 competent E. coil (Life technologies) on LB culture plates with ampicillin. .. The amplified plasmids were isolated, purified from bacterial cultures by HiSpeed Plasmid Midi Kit (Qiagen).

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Total genomic DNA (gDNA) of SA-01 was isolated using a modified SDS-proteinase K treatment method ( ) and restriction digested using the endonuclease Sau3AI (New England BioLabs, Beverly, MA), using serial dilutions of enzyme ranging from 250 U ml−1 to 4.6 U ml−1 (15 min at 37°C). .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin. .. The insertion of the TfRC1 gene into the plasmid was verified by performing PCR on the isolated plasmid using the forward and reverse primers (T7 forward and BGH reverse) provided with the pcDNA 3.1 Directional TOPO expression kit (Life Technologies).

    Purification:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: The final 203 bp amplicons, the desired promoter sequences, were gel purified. .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression.

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: Amplicons were cleaned up with the QIAquick PCR Purification Kit (Qiagen, Hilden Germany) then mixed 1∶1. .. The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA).

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: It was amplified by Phusion HF DNA polymerase (M0530L) and purified by the QIAquick PCR Purification Kit (#28106) and QIA gel Extraction kit (#28706). .. Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher).

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: The desired fragments were resolved in a 2% agarose gel (1× TAE buffer), purified, and ligated into a pCEP4 vector using T4 ligase in a 5:1 (insert/vector) ratio. .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    Article Title: Exome Sequencing Identifies a Novel CEACAM16 Mutation Associated with Autosomal Dominant Nonsyndromic Hearing Loss DFNA4B in a Chinese Family
    Article Snippet: Plasmid transfection, amplification and extraction The two kinds of plasmid pRK5-wild type (WT) and G169R mutant (MT) CEACAM16 -Flag were transformed into One Shot TOP10 competent E. coil (Life technologies) on LB culture plates with ampicillin. .. The amplified plasmids were isolated, purified from bacterial cultures by HiSpeed Plasmid Midi Kit (Qiagen).

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Size-fractionated DNA fragments were ligated into pTrueBlue at 1:2 and 1:3 vector/insert ratios, using an average 4.5-kb insert size for conversion of molar ratios to mass ratios, with T4 DNA ligase (New England BioLabs) (6 Weiss units) at 16°C for 12 h. Again, the plasmids were purified using a GFX PCR DNA and gel band purification kit. .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin. .. Plasmids were purified from these E. coli cells using the PureLink Quick Plasmid Miniprep kit (Life Technologies) as per manufacturer's instructions.

    Polymerase Chain Reaction:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: The hybridized DNA was column purified using a Qiagen PCR purification kit and the secondary nested PCR was completed. .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression.

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: PCR was performed on the amplicon mix using primer set F1 and IS1194. .. The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA).

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: The PCR product was cut with NEB CutSmart and ligated overnight at 16ºC to p3XFLAG-CMV™ -7.1 expression vector (E7533 SIGMA). .. Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher).

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: Cloning was performed using the TA cloning vector pJET1.2 (CloneJet PCR cloning kit; Fermentas, Glen Burnie, MD, USA). .. The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing.

    Article Title: α-Xylosidase plays essential roles in xyloglucan remodelling, maintenance of cell wall integrity, and seed germination in Arabidopsis thaliana
    Article Snippet: Transgenic plants The TRG1/XYL1 promoter region and the gene containing upstream and downstream regions were amplified from Ws genomic DNA by high-fidelity PCR with specific primers ( Supplementary Table S2 ) and PrimeSTAR DNA polymerase (Takara Bio Inc.). .. The product was cloned into the Gateway entry vector using the pENTR Directional TOPO Cloning Kit (Invitrogen, K2400-20SP) and transformed One Shot Chemically Competent Escherichia coli (Invitrogen, C4040-03).

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: To filter induced PCR mutations and distinguish possible allelic variants, at least four independent PCR products, and clones from each L1 transduction family member were capillary sequenced using 12 overlapping primer pairs ( ) distributed at ∼500-bp intervals covering the entire L1 sequence. .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Size-fractionated DNA fragments were ligated into pTrueBlue at 1:2 and 1:3 vector/insert ratios, using an average 4.5-kb insert size for conversion of molar ratios to mass ratios, with T4 DNA ligase (New England BioLabs) (6 Weiss units) at 16°C for 12 h. Again, the plasmids were purified using a GFX PCR DNA and gel band purification kit. .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations.

    Article Title: Fecal Components Modulate Human Astrovirus Infectivity in Cells and Reconstituted Intestinal Tissues
    Article Snippet: .. The PCR product was cloned into a pCR4-TOPO cloning vector (Invitrogen, USA), and the plasmid was transformed into chemically competent E. coli One Shot TOP10 cells (Invitrogen, USA). .. The plasmid was in vitro transcribed using a T7 RiboMAX large-scale RNA production system (Promega), and 10-fold dilution series (from 107 to 104 copies/ml) were used as a standard.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: The TfRC1 PCR product was directionally cloned into pcDNA 3.1 vector as per manufacturer's instructions (Life Technologies). .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin.

    Quantitative RT-PCR:

    Article Title: Fecal Components Modulate Human Astrovirus Infectivity in Cells and Reconstituted Intestinal Tissues
    Article Snippet: Paragraph title: Real-time RT-PCR quantification. ... The PCR product was cloned into a pCR4-TOPO cloning vector (Invitrogen, USA), and the plasmid was transformed into chemically competent E. coli One Shot TOP10 cells (Invitrogen, USA).

    De-Phosphorylation Assay:

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Digested DNA fragments ranging from 3 to 6 kbp were excised from the agarose gel and purified using a GFX PCR DNA and gel band purification kit (GE Healthcare, Buckinghamshire, United Kingdom). pTrueBlue was linearized using BamHI (New England Biolabs) for 3 h at 37°C (1 U μl−1 ), followed by dephosphorylation using Antarctic phosphatase (New England BioLabs) (0.93 U μl−1 ; 4 h at 37°C). .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations.

    CRISPR:

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: The resulting amplicon spanned the 3′ end of cas1 through the truncated IS1194 fragment with a repeat-s4-repeat CRISPR sandwiched in-between. .. The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA).

    Nested PCR:

    Article Title: Investigation of Changes in Tetracycline Repressor Binding upon Mutations in the Tetracycline Operator
    Article Snippet: Products of the nested PCR were used as the template for final amplification of the synthetic promoters using external primers that corresponded to the terminal 20 bp of each sequence. .. Each promoter was cloned into pGLOW-TOPO (Invitrogen 12567-020) upstream of the green fluorescence protein reporter gene (gfp ) and transformed into chemically competent Top10 (LacI–, TetR−) E. coli (Invitrogen, C404010) by heat shock at 42 °C for 45 s. Transformants were screened by GFP expression.

    Plasmid Preparation:

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA). .. To enable selection and replication in a lactococcal host, the pCR4Blunt-TOPO containing the desired insert (pS4TOPO) was fused to the lactococcal vector pGhost9 (pGh9). pS4TOPO and pGh9 were each cut with SpeI (Promega, WI USA).

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: The PCR product was cut with NEB CutSmart and ligated overnight at 16ºC to p3XFLAG-CMV™ -7.1 expression vector (E7533 SIGMA). .. Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher).

    Article Title: Does Wheat Genetically Modified for Disease Resistance Affect Root-Colonizing Pseudomonads and Arbuscular Mycorrhizal Fungi?
    Article Snippet: Cloning was performed using the TA cloning vector pJET1.2 (CloneJet PCR cloning kit; Fermentas, Glen Burnie, MD, USA). .. The constructs were transformed into chemically competent Escherichia coli One Shot® TOP 10 cells (Invitrogen, Carlsbad, CA, USA), and transformants containing the pJET1.2_pqqC construct were selected for sequencing.

    Article Title: α-Xylosidase plays essential roles in xyloglucan remodelling, maintenance of cell wall integrity, and seed germination in Arabidopsis thaliana
    Article Snippet: .. The product was cloned into the Gateway entry vector using the pENTR Directional TOPO Cloning Kit (Invitrogen, K2400-20SP) and transformed One Shot Chemically Competent Escherichia coli (Invitrogen, C4040-03). .. Kanamycin-resistant colonies were selected and plasmid DNA was prepared using the QIAprep Spin Miniprep Kit (Qiagen, 27104).

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: The desired fragments were resolved in a 2% agarose gel (1× TAE buffer), purified, and ligated into a pCEP4 vector using T4 ligase in a 5:1 (insert/vector) ratio. .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    Article Title: Exome Sequencing Identifies a Novel CEACAM16 Mutation Associated with Autosomal Dominant Nonsyndromic Hearing Loss DFNA4B in a Chinese Family
    Article Snippet: .. Plasmid transfection, amplification and extraction The two kinds of plasmid pRK5-wild type (WT) and G169R mutant (MT) CEACAM16 -Flag were transformed into One Shot TOP10 competent E. coil (Life technologies) on LB culture plates with ampicillin. ..

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Size-fractionated DNA fragments were ligated into pTrueBlue at 1:2 and 1:3 vector/insert ratios, using an average 4.5-kb insert size for conversion of molar ratios to mass ratios, with T4 DNA ligase (New England BioLabs) (6 Weiss units) at 16°C for 12 h. Again, the plasmids were purified using a GFX PCR DNA and gel band purification kit. .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations.

    Article Title: Fecal Components Modulate Human Astrovirus Infectivity in Cells and Reconstituted Intestinal Tissues
    Article Snippet: .. The PCR product was cloned into a pCR4-TOPO cloning vector (Invitrogen, USA), and the plasmid was transformed into chemically competent E. coli One Shot TOP10 cells (Invitrogen, USA). .. The plasmid was in vitro transcribed using a T7 RiboMAX large-scale RNA production system (Promega), and 10-fold dilution series (from 107 to 104 copies/ml) were used as a standard.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin. .. Plasmids were purified from these E. coli cells using the PureLink Quick Plasmid Miniprep kit (Life Technologies) as per manufacturer's instructions.

    Real-time Polymerase Chain Reaction:

    Article Title: Salt-responsive gut commensal modulates TH17 axis and disease
    Article Snippet: Paragraph title: DNA extraction from feces and qPCR of 16S rRNA genes ... Standard curves for quantification consisted in ten-fold serial dilutions in the range of 108 to 100 copies of E. coli (Invitrogen, C404010) or L. murinus isolate 16S rRNA gene amplified with primers 27F (5’-GTTTGATCCTGGCTCAG-3’) and 1492R (5’-CGGCTA CCTTGTTACGAC-3’) .

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher). .. The cell was post-transfected for 48 hours at 37°C, then the transfection efficacy was determined by qPCR quantification and western blot analysis.

    Selection:

    Article Title: Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis
    Article Snippet: The construct was first cloned into pCR4Blunt-TOPO (Invitrogen, CA, USA) and transformed into chemically competent One Shot TOP10 E. coli (Invitrogen, CA, USA). .. To enable selection and replication in a lactococcal host, the pCR4Blunt-TOPO containing the desired insert (pS4TOPO) was fused to the lactococcal vector pGhost9 (pGh9). pS4TOPO and pGh9 were each cut with SpeI (Promega, WI USA).

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations. .. Transformed cells were plated onto LB medium plates (100 μl plate−1 ) containing ampicillin (0.06 mg ml−1 ), isopropyl-β- d -thiogalactopyranoside (IPTG; 0.01 mg ml−1 ), and 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (0.04 mg ml−1 ) and incubated overnight at 37°C to verify DNA ligation using blue-white selection.

    Agarose Gel Electrophoresis:

    Article Title: Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation
    Article Snippet: The desired fragments were resolved in a 2% agarose gel (1× TAE buffer), purified, and ligated into a pCEP4 vector using T4 ligase in a 5:1 (insert/vector) ratio. .. Five microliters of the ligation product was used to transform One Shot TOP10 chemically competent bacteria (C404010; Invitrogen) as per the manufacturer’s instructions.

    Article Title: A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme
    Article Snippet: Digested DNA fragments ranging from 3 to 6 kbp were excised from the agarose gel and purified using a GFX PCR DNA and gel band purification kit (GE Healthcare, Buckinghamshire, United Kingdom). pTrueBlue was linearized using BamHI (New England Biolabs) for 3 h at 37°C (1 U μl−1 ), followed by dephosphorylation using Antarctic phosphatase (New England BioLabs) (0.93 U μl−1 ; 4 h at 37°C). .. Ligation mixtures were transformed into One Shot TOP10 E. coli competent cells (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations.

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: PCR product size was determined by electrophoresis at 50 V using a 0.7% agarose gel and visualized using SYBR Green Fluorescent dye. .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin.

    In Vitro:

    Article Title: Fecal Components Modulate Human Astrovirus Infectivity in Cells and Reconstituted Intestinal Tissues
    Article Snippet: The PCR product was cloned into a pCR4-TOPO cloning vector (Invitrogen, USA), and the plasmid was transformed into chemically competent E. coli One Shot TOP10 cells (Invitrogen, USA). .. The plasmid was in vitro transcribed using a T7 RiboMAX large-scale RNA production system (Promega), and 10-fold dilution series (from 107 to 104 copies/ml) were used as a standard.

    Electrophoresis:

    Article Title: Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells
    Article Snippet: PCR product size was determined by electrophoresis at 50 V using a 0.7% agarose gel and visualized using SYBR Green Fluorescent dye. .. The resulting plasmid was transformed into OneShot TOP10 E. coli competent cells (Life Technologies), which were then plated and grown at 37 °C overnight on LB agar plates containing 100 μg/mL of ampicillin.

    Transgenic Assay:

    Article Title: α-Xylosidase plays essential roles in xyloglucan remodelling, maintenance of cell wall integrity, and seed germination in Arabidopsis thaliana
    Article Snippet: Paragraph title: Transgenic plants ... The product was cloned into the Gateway entry vector using the pENTR Directional TOPO Cloning Kit (Invitrogen, K2400-20SP) and transformed One Shot Chemically Competent Escherichia coli (Invitrogen, C4040-03).

    Gel Extraction:

    Article Title: Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer
    Article Snippet: It was amplified by Phusion HF DNA polymerase (M0530L) and purified by the QIAquick PCR Purification Kit (#28106) and QIA gel Extraction kit (#28706). .. Transformation was performed with competent E. coli cells (One Shot Top10, #C404003, Thermofisher).

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