5 alpha electrocompetent e coli cells  (New England Biolabs)


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    New England Biolabs 5 alpha electrocompetent e coli cells
    5 Alpha Electrocompetent E Coli Cells, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5 alpha electrocompetent e coli cells/product/New England Biolabs
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    5 alpha electrocompetent e coli cells - by Bioz Stars, 2019-08
    93/100 stars

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    Related Articles

    Clone Assay:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: The PCR products were run on a gel to confirm amplicons are of the expected length. .. After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989). .. This ensuing library now contained the guide and donor sequences adjacent to the SNR52 promoter but was still missing the sgtail and an RNA polIII terminator.

    Centrifugation:

    Article Title: An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides
    Article Snippet: After clearing the cell debris by centrifugation at 30,000g for 15 min, the cell lysate was incubated with 0.5 ml fetuin beads at 4 °C for 1 h. The beads were then washed with 9 × 10 ml KBD buffer. .. The purified plasmid DNA was electroporated into NEB 5-alpha electrocompetent E. coli cells (NEB) for the next cycle of selection.

    Article Title: Isothermal folding of a light-up bio-orthogonal RNA origami nanoribbon
    Article Snippet: NEB 5-alpha electrocompetent E. coli cells (NEB) were diluted 1:1 with sterile milli-Q water in a prechilled tube: 20 μL of cells suspension were used for each electroporation experiment after the addition of 5 μL of labeled scaffold (DBS, noDBS1 or noDBS2). .. After electroporation, cells were allowed to recover for 25 min at 37 °C in pre-warm SOC.

    Amplification:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: The PCR products were run on a gel to confirm amplicons are of the expected length. .. After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989). .. This ensuing library now contained the guide and donor sequences adjacent to the SNR52 promoter but was still missing the sgtail and an RNA polIII terminator.

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol. .. From each pool, an aliquot was used to inoculate LB media for plasmid isolation (“prey” vectors).

    Article Title: Loss of a doublecortin (DCX)-domain protein causes structural defects in a tubulin-based organelle of Toxoplasma gondii and impairs host-cell invasion
    Article Snippet: After construction, plasmids were used to transform chemically competent TOP10 cells by heat shock or electrocompetent DH5α cells (NEB C2989) by electroporation. .. Converting from ptub-H2b-YFP to ptub-TgDCX-EGFP entailed removing the H2b sequence between the Nhe I and Bgl II sites and replacing it with the coding region of TgDCX extending from the second methionine to the amino acid before the stop codon, obtained by reverse transcription PCR (RT-PCR) using T. gondii total RNA and primer pair S5/AS5.

    Polymerase Chain Reaction:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: The PCR products were run on a gel to confirm amplicons are of the expected length. .. After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989). .. This ensuing library now contained the guide and donor sequences adjacent to the SNR52 promoter but was still missing the sgtail and an RNA polIII terminator.

    Article Title: Loss of a doublecortin (DCX)-domain protein causes structural defects in a tubulin-based organelle of Toxoplasma gondii and impairs host-cell invasion
    Article Snippet: After construction, plasmids were used to transform chemically competent TOP10 cells by heat shock or electrocompetent DH5α cells (NEB C2989) by electroporation. .. Plasmid DNA was isolated by standard procedures, and the constructions were verified by DNA sequencing. ptub-TgDCX-EGFP is a derivative of the plasmid ptub-H2b–yellow fluorescent protein (YFP; ).

    Real-time Polymerase Chain Reaction:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: The CustomArray-synthesized oligo library was diluted to 1 ng/μl and 1μl of the library was amplified with Kapa SYBR FAST qPCR Kit Master Mix (Kapa Biosystems) using unique primer pairs specific to each desired library (e.g. SGS1 tiling deletion, smORF library, etc.). .. After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989).

    Incubation:

    Article Title: An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides
    Article Snippet: After clearing the cell debris by centrifugation at 30,000g for 15 min, the cell lysate was incubated with 0.5 ml fetuin beads at 4 °C for 1 h. The beads were then washed with 9 × 10 ml KBD buffer. .. The purified plasmid DNA was electroporated into NEB 5-alpha electrocompetent E. coli cells (NEB) for the next cycle of selection.

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol. .. The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol.

    Modification:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: Primers used for oligo library amplification were further modified to contain the necessary overlaps to enable the library to be inserted into our vector backbone via Golden Gate cloning. .. After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989).

    Transformation Assay:

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: DNA from each pool was used for ligation into pKT25 [ ], which had been cut with SmaI and dephosphorylated by rAPid Alkaline Phosphatase (Roche, Mannheim, Germany). .. The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol. .. Ligation procedure was performed twice, resulting in ten transformations.

    Electroporation:

    Article Title: Isothermal folding of a light-up bio-orthogonal RNA origami nanoribbon
    Article Snippet: Electroporation and plug preparation has been done as previously described . .. NEB 5-alpha electrocompetent E. coli cells (NEB) were diluted 1:1 with sterile milli-Q water in a prechilled tube: 20 μL of cells suspension were used for each electroporation experiment after the addition of 5 μL of labeled scaffold (DBS, noDBS1 or noDBS2). .. The mixture of competent cells and labeled molecules was immediately transferred into a prechilled electroporation cuvette (0.2 cm gap cuvette, BioRad) and electroporated (Gene Pulser, BioRad; 1.8 kV, 200 Ω, 25 μF).

    Article Title: Loss of a doublecortin (DCX)-domain protein causes structural defects in a tubulin-based organelle of Toxoplasma gondii and impairs host-cell invasion
    Article Snippet: The final PCR products were gel-purified, cloned, and then sequenced as for the 5′RACE products. .. After construction, plasmids were used to transform chemically competent TOP10 cells by heat shock or electrocompetent DH5α cells (NEB C2989) by electroporation. .. Plasmid DNA was isolated by standard procedures, and the constructions were verified by DNA sequencing. ptub-TgDCX-EGFP is a derivative of the plasmid ptub-H2b–yellow fluorescent protein (YFP; ).

    Ligation:

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: DNA from each pool was used for ligation into pKT25 [ ], which had been cut with SmaI and dephosphorylated by rAPid Alkaline Phosphatase (Roche, Mannheim, Germany). .. The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol.

    Sequencing:

    Article Title: Loss of a doublecortin (DCX)-domain protein causes structural defects in a tubulin-based organelle of Toxoplasma gondii and impairs host-cell invasion
    Article Snippet: After construction, plasmids were used to transform chemically competent TOP10 cells by heat shock or electrocompetent DH5α cells (NEB C2989) by electroporation. .. Plasmid DNA was isolated by standard procedures, and the constructions were verified by DNA sequencing. ptub-TgDCX-EGFP is a derivative of the plasmid ptub-H2b–yellow fluorescent protein (YFP; ).

    Sonication:

    Article Title: An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides
    Article Snippet: The spheroplasts were lysed in 10 ml KBD buffer (10 mM Tris.HCl pH 7.4, 50 mM potassium glutamate, 3 mM DTT, 0.02% v/v Triton x-100, 10% glycerol, 0.1 mg ml−1 sonicated herring sperm DNA (Sigma). .. The purified plasmid DNA was electroporated into NEB 5-alpha electrocompetent E. coli cells (NEB) for the next cycle of selection.

    Isolation:

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: Five pools of DNA from 1.7 to 3 kbp were isolated, cleaned and treated with Klenow fragment to generate blunt ends. .. The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol.

    Subcloning:

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: Genomic DNA was sheared using the nebulizer from the TOPO Shotgun Subcloning Kit (Invitrogen, Carlsbad, CA, USA) and separated on a preparative agarose gel. .. The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol.

    Negative Control:

    Article Title: Isothermal folding of a light-up bio-orthogonal RNA origami nanoribbon
    Article Snippet: NEB 5-alpha electrocompetent E. coli cells (NEB) were diluted 1:1 with sterile milli-Q water in a prechilled tube: 20 μL of cells suspension were used for each electroporation experiment after the addition of 5 μL of labeled scaffold (DBS, noDBS1 or noDBS2). .. The mixture of competent cells and labeled molecules was immediately transferred into a prechilled electroporation cuvette (0.2 cm gap cuvette, BioRad) and electroporated (Gene Pulser, BioRad; 1.8 kV, 200 Ω, 25 μF).

    Labeling:

    Article Title: Isothermal folding of a light-up bio-orthogonal RNA origami nanoribbon
    Article Snippet: Electroporation and plug preparation has been done as previously described . .. NEB 5-alpha electrocompetent E. coli cells (NEB) were diluted 1:1 with sterile milli-Q water in a prechilled tube: 20 μL of cells suspension were used for each electroporation experiment after the addition of 5 μL of labeled scaffold (DBS, noDBS1 or noDBS2). .. The mixture of competent cells and labeled molecules was immediately transferred into a prechilled electroporation cuvette (0.2 cm gap cuvette, BioRad) and electroporated (Gene Pulser, BioRad; 1.8 kV, 200 Ω, 25 μF).

    Purification:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: The PCR products were run on a gel to confirm amplicons are of the expected length. .. After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989). .. This ensuing library now contained the guide and donor sequences adjacent to the SNR52 promoter but was still missing the sgtail and an RNA polIII terminator.

    Article Title: An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides
    Article Snippet: The plasmid DNA was then purified from the eluent according to the protocol of the Qiagen QIAprep Kit. .. The purified plasmid DNA was electroporated into NEB 5-alpha electrocompetent E. coli cells (NEB) for the next cycle of selection. .. HSA-depleted or IgG-depleted human serum was prepared by incubating human serum with Affi-Gel Blue beads (Bio-Rad) or Protein A beads (Thermo Fisher Scientific) according to the manufacturer's protocols, respectively.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Loss of a doublecortin (DCX)-domain protein causes structural defects in a tubulin-based organelle of Toxoplasma gondii and impairs host-cell invasion
    Article Snippet: After construction, plasmids were used to transform chemically competent TOP10 cells by heat shock or electrocompetent DH5α cells (NEB C2989) by electroporation. .. Plasmid DNA was isolated by standard procedures, and the constructions were verified by DNA sequencing. ptub-TgDCX-EGFP is a derivative of the plasmid ptub-H2b–yellow fluorescent protein (YFP; ).

    Plasmid Preparation:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: The PCR products were run on a gel to confirm amplicons are of the expected length. .. After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989). .. This ensuing library now contained the guide and donor sequences adjacent to the SNR52 promoter but was still missing the sgtail and an RNA polIII terminator.

    Article Title: An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides
    Article Snippet: The plasmid DNA was then purified from the eluent according to the protocol of the Qiagen QIAprep Kit. .. The purified plasmid DNA was electroporated into NEB 5-alpha electrocompetent E. coli cells (NEB) for the next cycle of selection. .. HSA-depleted or IgG-depleted human serum was prepared by incubating human serum with Affi-Gel Blue beads (Bio-Rad) or Protein A beads (Thermo Fisher Scientific) according to the manufacturer's protocols, respectively.

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol. .. Each transformation was plated on three LB/Kanamycin (50 µg/mL) plates (15 cm in diameter) and incubated overnight at 37 °C.

    Article Title: Loss of a doublecortin (DCX)-domain protein causes structural defects in a tubulin-based organelle of Toxoplasma gondii and impairs host-cell invasion
    Article Snippet: Paragraph title: Plasmid construction ... After construction, plasmids were used to transform chemically competent TOP10 cells by heat shock or electrocompetent DH5α cells (NEB C2989) by electroporation.

    Functional Assay:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989). .. After PCR purification (Zymo Research), the amplicon is cloned into the BsmBI-containing library vector (XG128) using a standard Golden Gate protocol with BsmBI (NEB R0580S) and T4 ligase (NEB M0202S) then electroporated into 5-alpha Electrocompetent E.coli cells (NEB C2989).

    Selection:

    Article Title: An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides
    Article Snippet: The plasmid DNA was then purified from the eluent according to the protocol of the Qiagen QIAprep Kit. .. The purified plasmid DNA was electroporated into NEB 5-alpha electrocompetent E. coli cells (NEB) for the next cycle of selection. .. HSA-depleted or IgG-depleted human serum was prepared by incubating human serum with Affi-Gel Blue beads (Bio-Rad) or Protein A beads (Thermo Fisher Scientific) according to the manufacturer's protocols, respectively.

    Agarose Gel Electrophoresis:

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: Genomic DNA was sheared using the nebulizer from the TOPO Shotgun Subcloning Kit (Invitrogen, Carlsbad, CA, USA) and separated on a preparative agarose gel. .. The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol.

    Lysis:

    Article Title: The Peptidoglycan Pattern of Staphylococcus carnosus TM300—Detailed Analysis and Variations Due to Genetic and Metabolic Influences
    Article Snippet: Chromosomal DNA from S. carnosus was isolated [ ] using lysostaphin (0.5 mg/mL) for cell lysis. .. The resulting plasmids were transformed into NEB 5-alpha electrocompetent E. coli cells (New England Biolabs, Ipswich, MA, USA) according to the manufacturer’s protocol.

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    NEB 5 alpha Electrocompetent E coli 6x0 1 ml
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    93
    New England Biolabs 5 alpha electrocompetent e coli cells
    5 Alpha Electrocompetent E Coli Cells, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5 alpha electrocompetent e coli cells/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    5 alpha electrocompetent e coli cells - by Bioz Stars, 2019-08
    93/100 stars
      Buy from Supplier

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