one shot top10  (Thermo Fisher)


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    Name:
    One Shot TOP10 Chemically Competent E coli
    Description:
    One Shot TOP10 Chemically Competent E coli are provided at a transformation efficiency of 1 x 109 cfu µg plasmid DNA and are ideal for high efficiency cloning and plasmid propagation They allow stable replication of high copy number plasmids and are the same competent cells that come with many of our cloning kits Features of One Shot TOP10 cells include • Maximization of cloning efficiency in a single tube format• Enhanced genomic DNA cloning capabilitiesEasy to use One Shot formatThe single tube single use format allows for all steps of the transformation protocol up to plating to take place in the same tube thereby helping to save time and prevent contamination Versatile cloning capabilitiesOne Shot TOP10 E coli cells are similar to the DH10B strain and offer the following features • hsdR for efficient transformation of unmethylated DNA from PCR amplifications• mcrA for efficient transformation of methylated DNA from genomic preparations• lacZΔM15 for blue white color screening of recombinant clones• endA1 for cleaner DNA preparations and better results in downstream applications due to elimination of nonspecific digestion by Endonuclease I• recA1 for reduced occurrence of nonspecific recombination in cloned DNAGenotype F mcrA Δ mrr hsdRMS mcrBC Φ80lacZΔM15 Δ lacX74 recA1 araD139 Δ araleu 7697 galU galK rpsL StrR endA1 nupGFind the strain and format that you needWe also offer many other different strains and formats of chemically competent cells and electrocompetent cells to help meet your specific transformation needs If you require high throughput transformation choose from our collection of MultiShot formatted comp cells
    Catalog Number:
    c404003
    Price:
    None
    Applications:
    Chemically Competent Cells for Cloning|Cloning|Transformation
    Category:
    Competent Cells Strains
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    Structured Review

    Thermo Fisher one shot top10
    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli <t>Top10</t> cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.
    One Shot TOP10 Chemically Competent E coli are provided at a transformation efficiency of 1 x 109 cfu µg plasmid DNA and are ideal for high efficiency cloning and plasmid propagation They allow stable replication of high copy number plasmids and are the same competent cells that come with many of our cloning kits Features of One Shot TOP10 cells include • Maximization of cloning efficiency in a single tube format• Enhanced genomic DNA cloning capabilitiesEasy to use One Shot formatThe single tube single use format allows for all steps of the transformation protocol up to plating to take place in the same tube thereby helping to save time and prevent contamination Versatile cloning capabilitiesOne Shot TOP10 E coli cells are similar to the DH10B strain and offer the following features • hsdR for efficient transformation of unmethylated DNA from PCR amplifications• mcrA for efficient transformation of methylated DNA from genomic preparations• lacZΔM15 for blue white color screening of recombinant clones• endA1 for cleaner DNA preparations and better results in downstream applications due to elimination of nonspecific digestion by Endonuclease I• recA1 for reduced occurrence of nonspecific recombination in cloned DNAGenotype F mcrA Δ mrr hsdRMS mcrBC Φ80lacZΔM15 Δ lacX74 recA1 araD139 Δ araleu 7697 galU galK rpsL StrR endA1 nupGFind the strain and format that you needWe also offer many other different strains and formats of chemically competent cells and electrocompetent cells to help meet your specific transformation needs If you require high throughput transformation choose from our collection of MultiShot formatted comp cells
    https://www.bioz.com/result/one shot top10/product/Thermo Fisher
    Average 99 stars, based on 62 article reviews
    Price from $9.99 to $1999.99
    one shot top10 - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach"

    Article Title: AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0137652

    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli Top10 cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.
    Figure Legend Snippet: AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli Top10 cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.

    Techniques Used: Clone Assay, Polymerase Chain Reaction, Amplification, Sequencing, Expressing, Plasmid Preparation, Transformation Assay, Derivative Assay, Incubation

    AQUA Expression—combined cloning and protein expression. (a) Timeline for AQUA Expression. Cloning and production of recombinant protein in E . coli may be performed within 24 h starting with the PCR until the bacteria are harvested the next day. (b) A 3-DNA fragment cloning was performed by inserting the coding sequence for the red fluorescent protein mCherry into a bacterial T7 promoter-driven expression vector. The vector was split into two parts within the resistance gene for the antibiotic spectinomycin. Therefore, only correctly assembled fragments allow cell growth. (c) AQUA Expression in the expression strain BL21 (DE3) results in red colored bacteria due to mCherry protein production, while the TOP10 strain—lacking the required T7 RNA polymerase—remains colorless.
    Figure Legend Snippet: AQUA Expression—combined cloning and protein expression. (a) Timeline for AQUA Expression. Cloning and production of recombinant protein in E . coli may be performed within 24 h starting with the PCR until the bacteria are harvested the next day. (b) A 3-DNA fragment cloning was performed by inserting the coding sequence for the red fluorescent protein mCherry into a bacterial T7 promoter-driven expression vector. The vector was split into two parts within the resistance gene for the antibiotic spectinomycin. Therefore, only correctly assembled fragments allow cell growth. (c) AQUA Expression in the expression strain BL21 (DE3) results in red colored bacteria due to mCherry protein production, while the TOP10 strain—lacking the required T7 RNA polymerase—remains colorless.

    Techniques Used: Expressing, Clone Assay, Recombinant, Polymerase Chain Reaction, Sequencing, Plasmid Preparation

    AQUA Cloning: a dvanced qu ick a ssembly cloning. (a) DNA parts are produced by PCR, or restriction digest (or both). Oligonucleotides are designed to contribute flanking homologous regions to adjacent DNA fragments of optimally 32 bp in length. DNA parts are assembled into a circular plasmid by sequence-determined directionality. (b) AQUA Cloning work-flow. (1) DNA parts are generated by PCR amplification, or derived from an enzymatic digestion. (2) Next, DNA parts are purified by gel-electrophoresis and (3) mixed and simply incubated in water prior to transformation into chemically competent E . coli Top10 cells for in vivo assembly. (4) Finally, obtained colonies are confirmed for correct assembly by standard methods such as analytical PCR, restriction digest, or comprehensive sequencing.
    Figure Legend Snippet: AQUA Cloning: a dvanced qu ick a ssembly cloning. (a) DNA parts are produced by PCR, or restriction digest (or both). Oligonucleotides are designed to contribute flanking homologous regions to adjacent DNA fragments of optimally 32 bp in length. DNA parts are assembled into a circular plasmid by sequence-determined directionality. (b) AQUA Cloning work-flow. (1) DNA parts are generated by PCR amplification, or derived from an enzymatic digestion. (2) Next, DNA parts are purified by gel-electrophoresis and (3) mixed and simply incubated in water prior to transformation into chemically competent E . coli Top10 cells for in vivo assembly. (4) Finally, obtained colonies are confirmed for correct assembly by standard methods such as analytical PCR, restriction digest, or comprehensive sequencing.

    Techniques Used: Clone Assay, Produced, Polymerase Chain Reaction, Plasmid Preparation, Sequencing, Flow Cytometry, Generated, Amplification, Derivative Assay, Purification, Nucleic Acid Electrophoresis, Incubation, Transformation Assay, In Vivo

    Related Articles

    Clone Assay:

    Article Title: Deep-sea corals near cold seeps associate with chemoautotrophic bacteria that are related to the symbionts of cold seep and hydrothermal vent mussels
    Article Snippet: .. Transformation was conducted using One Shot™ TOP10 Chemically Competent E. coli (Invitrogen, USA) by combining 3µl of TOPO cloning reaction with 50 µL of E. coli cells. ..

    Article Title: Copper nanoparticles prompt the activity of estuarine denitrifying bacterial communities at relevant environmental concentrations
    Article Snippet: .. PCR products were cloned using the TOPO-TA cloning kit with pCR 2.1-TOPO and One Shot TOP10 Chemically Competent E. coli (Invitrogen – Thermo Fisher). .. Colonies developed for 10 h in solid medium (40 g L-1 LB Broth, ampicillin 50 μg ml-1 and X-gal 100 mmol L-1 ) at 37 °C, and then selected plasmid-inserted colonies were grown overnight in liquid medium.

    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: A novel mycovirus evokes transcriptional rewiring in the fungus Malassezia and stimulates interferon-β production in macrophages
    Article Snippet: .. Next, the adapter-ligated/RT primer mixture was used as template with the AffinityScript cDNA Synthesis Kit (Agilent, 600559) with the following reaction conditions: 25°C for 5 minutes, 55°C for 15 minutes, 95°C for 5 minutes. cDNA was PCR amplified using the RT primer, cloned into the pCR-Blunt vector using the Zero Blunt PCR Cloning Kit (Invitrogen, K270020), and transformed into E. coli with One Shot TOP10 Chemically Competent E. coli (Invitrogen, C404003). .. Plasmids were extracted from purified bacterial colonies and subjected to Sanger sequencing (Genewiz) using universal M13F and M13R primers.

    Amplification:

    Article Title: A novel mycovirus evokes transcriptional rewiring in the fungus Malassezia and stimulates interferon-β production in macrophages
    Article Snippet: .. Next, the adapter-ligated/RT primer mixture was used as template with the AffinityScript cDNA Synthesis Kit (Agilent, 600559) with the following reaction conditions: 25°C for 5 minutes, 55°C for 15 minutes, 95°C for 5 minutes. cDNA was PCR amplified using the RT primer, cloned into the pCR-Blunt vector using the Zero Blunt PCR Cloning Kit (Invitrogen, K270020), and transformed into E. coli with One Shot TOP10 Chemically Competent E. coli (Invitrogen, C404003). .. Plasmids were extracted from purified bacterial colonies and subjected to Sanger sequencing (Genewiz) using universal M13F and M13R primers.

    Sample Prep:

    Article Title: In vivo analysis of the Escherichia coli ultrastructure by small-angle scattering
    Article Snippet: .. Sample preparation One Shot TOP10 chemically competent E. coli cells from Invitrogen (K-12 strain, similar to the DH10B strain) were used in this study. .. Colonies were grown in LB medium (Sigma–Aldrich) with ampicillin (100 µg ml−1 , Euromedex) at 37°C to an OD600 of ∼1 (∼8 × 108 cells ml−1 ).

    Generated:

    Article Title: SNAIL is a key regulator of alveolar rhabdomyosarcoma tumor growth and differentiation through repression of MYF5 and MYOD function
    Article Snippet: .. Bacterial transformations of One Shot™ TOP10 Chemically Competent E. coli (Invitrogen, USA) with generated UBC@pNL and MYF5@pNL plasmids were performed and LB Agar plates with 100 µg/ml ampicillin were used for bacterial selection. .. 6 clones of each plasmid were purified from selected bacterial colonies using QIAfilter Plasmid Maxi Kit (Qiagen, Germany).

    Selection:

    Article Title: SNAIL is a key regulator of alveolar rhabdomyosarcoma tumor growth and differentiation through repression of MYF5 and MYOD function
    Article Snippet: .. Bacterial transformations of One Shot™ TOP10 Chemically Competent E. coli (Invitrogen, USA) with generated UBC@pNL and MYF5@pNL plasmids were performed and LB Agar plates with 100 µg/ml ampicillin were used for bacterial selection. .. 6 clones of each plasmid were purified from selected bacterial colonies using QIAfilter Plasmid Maxi Kit (Qiagen, Germany).

    Article Title: An improved Nicotiana benthamiana strain for aphid and whitefly research
    Article Snippet: .. The reactions were placed in thermal cycler at 50°C for 1 h. Then, 2 μl of the reaction were transformed into 50 μl of the One Shot(tm) Top10 chemically competent cells (Invitrogen, www.thermofisher.com ) and plated with 50 μl/ml kanamycin selection. ..

    Polymerase Chain Reaction:

    Article Title: Copper nanoparticles prompt the activity of estuarine denitrifying bacterial communities at relevant environmental concentrations
    Article Snippet: .. PCR products were cloned using the TOPO-TA cloning kit with pCR 2.1-TOPO and One Shot TOP10 Chemically Competent E. coli (Invitrogen – Thermo Fisher). .. Colonies developed for 10 h in solid medium (40 g L-1 LB Broth, ampicillin 50 μg ml-1 and X-gal 100 mmol L-1 ) at 37 °C, and then selected plasmid-inserted colonies were grown overnight in liquid medium.

    Article Title: A novel mycovirus evokes transcriptional rewiring in the fungus Malassezia and stimulates interferon-β production in macrophages
    Article Snippet: .. Next, the adapter-ligated/RT primer mixture was used as template with the AffinityScript cDNA Synthesis Kit (Agilent, 600559) with the following reaction conditions: 25°C for 5 minutes, 55°C for 15 minutes, 95°C for 5 minutes. cDNA was PCR amplified using the RT primer, cloned into the pCR-Blunt vector using the Zero Blunt PCR Cloning Kit (Invitrogen, K270020), and transformed into E. coli with One Shot TOP10 Chemically Competent E. coli (Invitrogen, C404003). .. Plasmids were extracted from purified bacterial colonies and subjected to Sanger sequencing (Genewiz) using universal M13F and M13R primers.

    Transformation Assay:

    Article Title: Deep-sea corals near cold seeps associate with chemoautotrophic bacteria that are related to the symbionts of cold seep and hydrothermal vent mussels
    Article Snippet: .. Transformation was conducted using One Shot™ TOP10 Chemically Competent E. coli (Invitrogen, USA) by combining 3µl of TOPO cloning reaction with 50 µL of E. coli cells. ..

    Article Title: A clinically-relevant polymorphism in the Na+/taurocholate cotransporting polypeptide (NTCP) occurs at a rheostat position
    Article Snippet: .. Mutated cDNA was then transformed into One Shot TOP10 cells (Thermo Fisher Scientific, Waltham, MA) and bacterial colonies were randomly selected. .. The cDNA was isolated using QIAGEN (Hilden, Germany) mini prep kits and sequenced (GENEWIZ, South Plainfield, NJ).

    Article Title: An improved Nicotiana benthamiana strain for aphid and whitefly research
    Article Snippet: .. The reactions were placed in thermal cycler at 50°C for 1 h. Then, 2 μl of the reaction were transformed into 50 μl of the One Shot(tm) Top10 chemically competent cells (Invitrogen, www.thermofisher.com ) and plated with 50 μl/ml kanamycin selection. ..

    Article Title: A novel mycovirus evokes transcriptional rewiring in the fungus Malassezia and stimulates interferon-β production in macrophages
    Article Snippet: .. Next, the adapter-ligated/RT primer mixture was used as template with the AffinityScript cDNA Synthesis Kit (Agilent, 600559) with the following reaction conditions: 25°C for 5 minutes, 55°C for 15 minutes, 95°C for 5 minutes. cDNA was PCR amplified using the RT primer, cloned into the pCR-Blunt vector using the Zero Blunt PCR Cloning Kit (Invitrogen, K270020), and transformed into E. coli with One Shot TOP10 Chemically Competent E. coli (Invitrogen, C404003). .. Plasmids were extracted from purified bacterial colonies and subjected to Sanger sequencing (Genewiz) using universal M13F and M13R primers.

    Plasmid Preparation:

    Article Title: A novel mycovirus evokes transcriptional rewiring in the fungus Malassezia and stimulates interferon-β production in macrophages
    Article Snippet: .. Next, the adapter-ligated/RT primer mixture was used as template with the AffinityScript cDNA Synthesis Kit (Agilent, 600559) with the following reaction conditions: 25°C for 5 minutes, 55°C for 15 minutes, 95°C for 5 minutes. cDNA was PCR amplified using the RT primer, cloned into the pCR-Blunt vector using the Zero Blunt PCR Cloning Kit (Invitrogen, K270020), and transformed into E. coli with One Shot TOP10 Chemically Competent E. coli (Invitrogen, C404003). .. Plasmids were extracted from purified bacterial colonies and subjected to Sanger sequencing (Genewiz) using universal M13F and M13R primers.

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