electrocomp kit  (Thermo Fisher)


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    Name:
    TOP10 Electrocomp Kit
    Description:
    TOP10 E coli are provided at a transformation efficiency of 1 × 1010 cfu µg supercoiled DNA and are ideal for high efficiency cloning and plasmid propagation They allow stable replication of high copy number plasmids The genotype of TOP10 cells is similar to the DH10B strain and offers 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 preparations of DNA and better results in downstream applications due to the elimination of non specific digestion by Endonuclease I• recA1 for reduced occurrence of non specific recombination in cloned DNAOne Shot TOP10 Kits are available with either chemically competent or electrocompetent E coli to fit your specific transformation needs TOP10 E coli are also available in the high throughput MultiShot format Note This kit contains enough electrocompetent cells for 25 x 40 µL rxns or 50 x 20 µL rxns Please see the product manual for reaction scale recommendations Genotype F mcrA Δ mrr hsdRMS mcrBC φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ ara leu 7697 galU galK rpsL endA1 nupG
    Catalog Number:
    c66411
    Price:
    None
    Applications:
    Cloning|Electrocompetent Cells for Cloning|Transformation
    Category:
    Competent Cells Strains
    Buy from Supplier


    Structured Review

    Thermo Fisher electrocomp kit
    TOP10 E coli are provided at a transformation efficiency of 1 × 1010 cfu µg supercoiled DNA and are ideal for high efficiency cloning and plasmid propagation They allow stable replication of high copy number plasmids The genotype of TOP10 cells is similar to the DH10B strain and offers 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 preparations of DNA and better results in downstream applications due to the elimination of non specific digestion by Endonuclease I• recA1 for reduced occurrence of non specific recombination in cloned DNAOne Shot TOP10 Kits are available with either chemically competent or electrocompetent E coli to fit your specific transformation needs TOP10 E coli are also available in the high throughput MultiShot format Note This kit contains enough electrocompetent cells for 25 x 40 µL rxns or 50 x 20 µL rxns Please see the product manual for reaction scale recommendations Genotype F mcrA Δ mrr hsdRMS mcrBC φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ ara leu 7697 galU galK rpsL endA1 nupG
    https://www.bioz.com/result/electrocomp kit/product/Thermo Fisher
    Average 90 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    electrocomp kit - by Bioz Stars, 2020-04
    90/100 stars

    Related Products / Commonly Used Together

    electrocompetent e. coli top10f
    electroporation
    top10f

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    Related Articles

    Clone Assay:

    Article Title: Quantification of the physiochemical constraints on the export of spider silk proteins by Salmonella type III secretion
    Article Snippet: .. All cloning was conducted using TOP10 E. coli (Invitrogen, #C664-11). .. Three medias were used: LB Miller (Difco, #244510), L Broth (Difco, #241210) and SPI-1 Inducing Media (IM) (LB, 0.3 M NaCl [ ]).

    Article Title: Fis Regulates Transcriptional Induction of RpoS in Salmonella enterica
    Article Snippet: .. RACE reaction products were eluted from 1% agarose gels by using the NucleoTrap gel extraction kit (BD Biosciences), cloned into a pCR4-TOPO vector (Invitrogen) and subsequently used to transform TOP 10 Electrocomp cells (Invitrogen). ..

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Gene coverage and ORF enrichment was analyzed by sequencing of individual E. coli XL1 blue MRF' clones infected with TcdB-gene fragment phage.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The amplicon was blunt-end cloned using T4 DNA ligase and ligase buffer (ThermoFisher #EL0011). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The amplicon was blunt-end cloned using T4 DNA ligase and ligase buffer (ThermoFisher #EL0011). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Gene coverage and ORF enrichment was analyzed by sequencing of individual E. coli XL1 blue MRF' clones infected with TcdB-gene fragment phage.

    Amplification:

    Article Title: Fis Regulates Transcriptional Induction of RpoS in Salmonella enterica
    Article Snippet: RNA served as the template for the 5′-rapid-amplification-of-cDNA-end (5′RACE) reactions performed with a BD SMART RACE cDNA amplification kit as described by the manufacturer (BD Biosciences) with one exception. .. RACE reaction products were eluted from 1% agarose gels by using the NucleoTrap gel extraction kit (BD Biosciences), cloned into a pCR4-TOPO vector (Invitrogen) and subsequently used to transform TOP 10 Electrocomp cells (Invitrogen).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The amplicon was blunt-end cloned using T4 DNA ligase and ligase buffer (ThermoFisher #EL0011). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The amplicon was blunt-end cloned using T4 DNA ligase and ligase buffer (ThermoFisher #EL0011). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Agarose Gel Electrophoresis:

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The PCR product was run on an agarose gel and purified with the Gel-Extraction Kit (Qiagen). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The PCR product was run on an agarose gel and purified with the Gel-Extraction Kit (Qiagen). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Fluorescence:

    Article Title: Quantification of the physiochemical constraints on the export of spider silk proteins by Salmonella type III secretion
    Article Snippet: Microorganisms and Media Salmonella typhimurium SL1344 was used for all secretion and fluorescence experiments. .. All cloning was conducted using TOP10 E. coli (Invitrogen, #C664-11).

    Ligation:

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml). .. The second library was constructed similarly to the first library, except that 8 instead of 20 cycles were run on the error-prone PCR product with primers containing restriction enzyme tails.

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: .. Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Successfully transformed TOP10F' cells were selected on 2 × YT agar [1.6% (w/v) tryptone, 1% (w/v) yeast extract, 0.05% (w/v) NaCl, 1.2% (w/v) agar] supplemented with 100 mM glucose and 100 μg/mL ampicillin.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Ligation solution was purified with PCR cleanup (A & A Technologies). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Ligation solution was purified with PCR cleanup (A & A Technologies). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml). .. The second library was constructed similarly to the first library, except that 8 instead of 20 cycles were run on the error-prone PCR product with primers containing restriction enzyme tails.

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: .. Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Successfully transformed TOP10F' cells were selected on 2 × YT agar [1.6% (w/v) tryptone, 1% (w/v) yeast extract, 0.05% (w/v) NaCl, 1.2% (w/v) agar] supplemented with 100 mM glucose and 100 μg/mL ampicillin.

    Mutagenesis:

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Paragraph title: Construction of the bla CTX-M-15 mutant library ... The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455). .. The double mutant N135D-G239S was constructed using bla CTX-M-15 N135D as template.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455). .. The double mutant N135D-G239S was constructed using bla CTX-M-15 N135D as template.

    Isolation:

    Article Title: Fis Regulates Transcriptional Induction of RpoS in Salmonella enterica
    Article Snippet: RNA was isolated from wild-type LT2 cells during exponential phase by using an RNeasy minikit (Qiagen). .. RACE reaction products were eluted from 1% agarose gels by using the NucleoTrap gel extraction kit (BD Biosciences), cloned into a pCR4-TOPO vector (Invitrogen) and subsequently used to transform TOP 10 Electrocomp cells (Invitrogen).

    Construct:

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml). .. The second library was constructed similarly to the first library, except that 8 instead of 20 cycles were run on the error-prone PCR product with primers containing restriction enzyme tails.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455). .. The double mutants S133G-G239S and S133G-N135D were constructed using bla CTX-M-15 S133G as template DNA for PCR with primers introducing the mutations G239S and N135D, respectively.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455). .. The double mutants S133G-G239S and S133G-N135D were constructed using bla CTX-M-15 S133G as template DNA for PCR with primers introducing the mutations G239S and N135D, respectively.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml). .. The second library was constructed similarly to the first library, except that 8 instead of 20 cycles were run on the error-prone PCR product with primers containing restriction enzyme tails.

    Purification:

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Both products were cut (ThermoFisher enzymes: #ER1891 and #ER1341) and purified using QIAquick PCR purification kit (Qiagen), and dephosporylated using 1 µl Phosphatase fast digest (ThermoFisher #EF0654) and purified with QIAquick PCR purification (Qiagen). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: DNA product was purified using HiYield® Gel/PCR DNA fragment extraction kit, (SLG). .. Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Ligation solution was purified with PCR cleanup (A & A Technologies). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Ligation solution was purified with PCR cleanup (A & A Technologies). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Both products were cut (ThermoFisher enzymes: #ER1891 and #ER1341) and purified using QIAquick PCR purification kit (Qiagen), and dephosporylated using 1 µl Phosphatase fast digest (ThermoFisher #EF0654) and purified with QIAquick PCR purification (Qiagen). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: DNA product was purified using HiYield® Gel/PCR DNA fragment extraction kit, (SLG). .. Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad).

    Polymerase Chain Reaction:

    Article Title: Fis Regulates Transcriptional Induction of RpoS in Salmonella enterica
    Article Snippet: RACE reaction products were eluted from 1% agarose gels by using the NucleoTrap gel extraction kit (BD Biosciences), cloned into a pCR4-TOPO vector (Invitrogen) and subsequently used to transform TOP 10 Electrocomp cells (Invitrogen). .. Plasmids were analyzed by PCR amplification of the insert, followed by DNA sequencing.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Both products were cut (ThermoFisher enzymes: #ER1891 and #ER1341) and purified using QIAquick PCR purification kit (Qiagen), and dephosporylated using 1 µl Phosphatase fast digest (ThermoFisher #EF0654) and purified with QIAquick PCR purification (Qiagen). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Ligation solution was purified with PCR cleanup (A & A Technologies). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Ligation solution was purified with PCR cleanup (A & A Technologies). .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Both products were cut (ThermoFisher enzymes: #ER1891 and #ER1341) and purified using QIAquick PCR purification kit (Qiagen), and dephosporylated using 1 µl Phosphatase fast digest (ThermoFisher #EF0654) and purified with QIAquick PCR purification (Qiagen). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Infection:

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Gene coverage and ORF enrichment was analyzed by sequencing of individual E. coli XL1 blue MRF' clones infected with TcdB-gene fragment phage.

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Gene coverage and ORF enrichment was analyzed by sequencing of individual E. coli XL1 blue MRF' clones infected with TcdB-gene fragment phage.

    Gel Extraction:

    Article Title: Fis Regulates Transcriptional Induction of RpoS in Salmonella enterica
    Article Snippet: .. RACE reaction products were eluted from 1% agarose gels by using the NucleoTrap gel extraction kit (BD Biosciences), cloned into a pCR4-TOPO vector (Invitrogen) and subsequently used to transform TOP 10 Electrocomp cells (Invitrogen). ..

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The PCR product was run on an agarose gel and purified with the Gel-Extraction Kit (Qiagen). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The PCR product was run on an agarose gel and purified with the Gel-Extraction Kit (Qiagen). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Sequencing:

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml). .. The complete sequence of the bla CTX-M-15 gene and the β-lactamase promoter can be found in Supplementary Data .

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Gene coverage and ORF enrichment was analyzed by sequencing of individual E. coli XL1 blue MRF' clones infected with TcdB-gene fragment phage.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455). .. Mutations were verified with Sanger sequencing.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455). .. Mutations were verified with Sanger sequencing.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml). .. The complete sequence of the bla CTX-M-15 gene and the β-lactamase promoter can be found in Supplementary Data .

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Gene coverage and ORF enrichment was analyzed by sequencing of individual E. coli XL1 blue MRF' clones infected with TcdB-gene fragment phage.

    Electroporation:

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: .. Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Successfully transformed TOP10F' cells were selected on 2 × YT agar [1.6% (w/v) tryptone, 1% (w/v) yeast extract, 0.05% (w/v) NaCl, 1.2% (w/v) agar] supplemented with 100 mM glucose and 100 μg/mL ampicillin.

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: .. Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Successfully transformed TOP10F' cells were selected on 2 × YT agar [1.6% (w/v) tryptone, 1% (w/v) yeast extract, 0.05% (w/v) NaCl, 1.2% (w/v) agar] supplemented with 100 mM glucose and 100 μg/mL ampicillin.

    Transformation Assay:

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml). .. The second library was constructed similarly to the first library, except that 8 instead of 20 cycles were run on the error-prone PCR product with primers containing restriction enzyme tails.

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Successfully transformed TOP10F' cells were selected on 2 × YT agar [1.6% (w/v) tryptone, 1% (w/v) yeast extract, 0.05% (w/v) NaCl, 1.2% (w/v) agar] supplemented with 100 mM glucose and 100 μg/mL ampicillin.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455). .. Mutations were verified with Sanger sequencing.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: .. Each of the resulting plasmids pZS3-CTX-M-15N135D , pZS3-CTX-M-15S133G and pZS3-CTX-M-15G239S were transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455). .. Mutations were verified with Sanger sequencing.

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml). .. The second library was constructed similarly to the first library, except that 8 instead of 20 cycles were run on the error-prone PCR product with primers containing restriction enzyme tails.

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad). .. Successfully transformed TOP10F' cells were selected on 2 × YT agar [1.6% (w/v) tryptone, 1% (w/v) yeast extract, 0.05% (w/v) NaCl, 1.2% (w/v) agar] supplemented with 100 mM glucose and 100 μg/mL ampicillin.

    Plasmid Preparation:

    Article Title: Fis Regulates Transcriptional Induction of RpoS in Salmonella enterica
    Article Snippet: .. RACE reaction products were eluted from 1% agarose gels by using the NucleoTrap gel extraction kit (BD Biosciences), cloned into a pCR4-TOPO vector (Invitrogen) and subsequently used to transform TOP 10 Electrocomp cells (Invitrogen). ..

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Ligation was performed using vector:insert ratio of 1:10 and ligated using T4 DNA ligase and ligase buffer (ThermoFisher #EL0011). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: A 10-fold molar excess of gene fragments was ligated into PmeI (NEB) linearized and CIP (Calf Instestine Phosphatase, NEB) dephosphorylated pHORF3 library vector (Kügler et al., ) (16 h at 16°C, T4 DNA Ligase, Promega). .. Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad).

    Article Title: Collateral sensitivity constrains resistance evolution of the CTX-M-15 β-lactamase
    Article Snippet: Ligation was performed using vector:insert ratio of 1:10 and ligated using T4 DNA ligase and ligase buffer (ThermoFisher #EL0011). .. The product resulting from this ligation, containing the bla CTX-M-15 SNP library named pZS3-CTX-M-15-Mut2Vol2, was transformed into electrocompetent E. coli TOP10 (ThermoFisher #C66455) and selected on kanamycin (50 µg/ml).

    Article Title: Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile
    Article Snippet: A 10-fold molar excess of gene fragments was ligated into PmeI (NEB) linearized and CIP (Calf Instestine Phosphatase, NEB) dephosphorylated pHORF3 library vector (Kügler et al., ) (16 h at 16°C, T4 DNA Ligase, Promega). .. Five microliter of ligation was used to transform 25 μL of electrocompetent E. coli TOP10F' (TOP10F' Electrocomp™ Kit, Life Technologies) by electroporation (1.8 kV, MicroPulser™, BioRad).

    DNA Sequencing:

    Article Title: Fis Regulates Transcriptional Induction of RpoS in Salmonella enterica
    Article Snippet: RACE reaction products were eluted from 1% agarose gels by using the NucleoTrap gel extraction kit (BD Biosciences), cloned into a pCR4-TOPO vector (Invitrogen) and subsequently used to transform TOP 10 Electrocomp cells (Invitrogen). .. Plasmids were analyzed by PCR amplification of the insert, followed by DNA sequencing.

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    Thermo Fisher top10 electrocomp kit
    Top10 Electrocomp Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    top10 electrocomp kit - by Bioz Stars, 2020-04
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