neb turbo competent e coli high efficiency  (New England Biolabs)


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    Name:
    NEB Turbo Competent E coli High Efficiency
    Description:
    NEB Turbo Competent E coli High Efficiency 20x0 05 ml
    Catalog Number:
    c2984h
    Price:
    263
    Size:
    1 ml
    Category:
    Competent Bacteria
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    Structured Review

    New England Biolabs neb turbo competent e coli high efficiency
    NEB Turbo Competent E coli High Efficiency
    NEB Turbo Competent E coli High Efficiency 20x0 05 ml
    https://www.bioz.com/result/neb turbo competent e coli high efficiency/product/New England Biolabs
    Average 95 stars, based on 32 article reviews
    Price from $9.99 to $1999.99
    neb turbo competent e coli high efficiency - by Bioz Stars, 2020-02
    95/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: C-terminal mCherry fusions with a (GGGS)2 linker were generated by Gibson cloning. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: CRISPR-Cas9-Based Knockout of the Prion Protein and Its Effect on the Proteome
    Article Snippet: The respective oligonucleotide pairs were obtained from Life Technologies (Burlington, ON, Canada) and were customized to include overhangs compatible for ligation into MLM3636 linearized by digestion with BsmB1 (cat. no. R0580S; New England BioLabs, Ipswich, MA, USA), a cloning site located in this vector on the 3′ side of a U6 promoter element. .. Oligonucleotides were phosphorylated with polynucleotide kinase (cat. no. EK0031; Fermentas, Ottawa, ON, Canada), annealed and inserted into the gRNA plasmid using T4 DNA ligase (cat no. M0202S; New England Biolabs) and transformed into Turbo competent E. coli (cat no. C2984H; New England Biolabs).

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells. .. All templates were cloned into a T7 expression plasmid system—the PURExpress control vector from New England Biolabs, called pNP1 in the text, pJL1, or pCOLADuet-1 (71406-3, Novagen)—using Gibson assembly ( ).

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells. .. All templates were cloned into a T7 expression plasmid system—the PURExpress control vector from New England Biolabs, called pNP1 in the text, pJL1, or pCOLADuet-1 (71406-3, Novagen)—using Gibson assembly ( ).

    Article Title: Engineering a Model Cell for Rational Tuning of GPCR Signaling
    Article Snippet: .. NEB® Turbo Competent E. coli was used for all cloning experiments. .. Selection and growth of E. coli was performed in Lysogeny Broth (LB) medium at 37°C with aeration.

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: C-terminal mCherry fusions with a (GGGS)2 linker were generated by Gibson cloning. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Neq SSB-like protein Thermostable SSB from Nanoarchaeum equitans (Neq SSB-like protein) was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET29a(+) and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express Competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 30 °C, induced with 0.5 mM IPTG, expressed at 16 °C overnight (14 hr), and purified by IMAC FPLC using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences).

    Amplification:

    Article Title: Plasmid-normalized quantification of relative mitochondrial DNA copy number
    Article Snippet: The distance between both inserts was chosen to be greater than 500 bp to avoid the amplification of both targets as one amplicon in the qPCR reaction (Fig. ). .. Plasmids were extracted from NEB Turbo Competent E. coli (New England Biolabs, Ipswich, MA, USA) with PureYield Plasmid Midiprep kit (Promega Co., Madison, WI, USA) and the concentration was measured using Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA).

    Article Title: Regulation of Phosphoribosyl-Linked Serine Ubiquitination by Deubiquitinases DupA and DupB
    Article Snippet: NEB Turbo (NEB, C2984H). .. NEB Turbo cells were used for plasmid amplification and molecular cloning.

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Amplified cDNA and vector were subjected to electrophoresis on 1% agarose gels and extracted using a QIAquick Gel Extraction kit (Qiagen 28706). .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    DNA Ligation:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Vector and insert were ligated using the Rapid DNA Ligation kit (Roche Applied Science 11635379001) as per the manufacturer’s recommended protocol. .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Synthesized:

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: General template design and preparation DNA sequences encoding eforRed, dTomato, mOrange, ATF1, Ecarin, and Trx-Bx (batroxobin fused with thioredoxin as a solubility domain) genes were derived from the literature, codon-optimized for Escherichia coli , and synthesized as gBlocks or oligonucleotides by Integrated DNA Technologies. pPROEX-Aquamarine was a gift from F. Merola (plasmid #42889, Addgene), and pET29-sortaseA-penta-mutant was a gift from L. Griffith. .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells.

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: DNA sequences encoding eforRed, dTomato, mOrange, ATF1, Ecarin, and Trx-Bx (batroxobin fused with thioredoxin as a solubility domain) genes were derived from the literature, codon-optimized for Escherichia coli , and synthesized as gBlocks or oligonucleotides by Integrated DNA Technologies. pPROEX-Aquamarine was a gift from F. Merola (plasmid #42889, Addgene), and pET29-sortaseA-penta-mutant was a gift from L. Griffith. .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells.

    TA Cloning:

    Article Title: Plasmid-normalized quantification of relative mitochondrial DNA copy number
    Article Snippet: Two sequential TA cloning reactions were performed to insert the two targets into the pGEM-T vector (Promega Co., Madison, WI, USA). .. Plasmids were extracted from NEB Turbo Competent E. coli (New England Biolabs, Ipswich, MA, USA) with PureYield Plasmid Midiprep kit (Promega Co., Madison, WI, USA) and the concentration was measured using Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA).

    Construct:

    Article Title: Plasmid-normalized quantification of relative mitochondrial DNA copy number
    Article Snippet: In order to verify whether the two fluorescent dyes present different intensities of fluorescence emission and to normalize for differences between runs (master mix performance, instrument calibration, environmental variability, etc.) a plasmid construct was designed. .. Plasmids were extracted from NEB Turbo Competent E. coli (New England Biolabs, Ipswich, MA, USA) with PureYield Plasmid Midiprep kit (Promega Co., Madison, WI, USA) and the concentration was measured using Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA).

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo). .. Mutants were generated by QuikChange site-directed mutagenesis.

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo). .. Mutants were generated by QuikChange site-directed mutagenesis.

    Real-time Polymerase Chain Reaction:

    Article Title: Plasmid-normalized quantification of relative mitochondrial DNA copy number
    Article Snippet: The distance between both inserts was chosen to be greater than 500 bp to avoid the amplification of both targets as one amplicon in the qPCR reaction (Fig. ). .. Plasmids were extracted from NEB Turbo Competent E. coli (New England Biolabs, Ipswich, MA, USA) with PureYield Plasmid Midiprep kit (Promega Co., Madison, WI, USA) and the concentration was measured using Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA).

    Incubation:

    Article Title: Rpn (YhgA-Like) Proteins of Escherichia coli K-12 and Their Contribution to RecA-Independent Horizontal Transfer
    Article Snippet: Plasmids were prepared from E. coli Turbo cells (catalog number C2984; New England BioLabs [NEB]). .. For temperature-sensitive plasmids, incubation at 30°C was used to permit plasmid replication and incubation at 42°C was used to remove the plasmid.

    Gel Extraction:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Amplified cDNA and vector were subjected to electrophoresis on 1% agarose gels and extracted using a QIAquick Gel Extraction kit (Qiagen 28706). .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Activity Assay:

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Endonuclease activity on supercoiled plasmids was verified according to previously published methods [ ] (see Supporting information ).

    Expressing:

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: Transgenes were cloned into a pET21/28-derived bacterial expression vector. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: Regulation of Phosphoribosyl-Linked Serine Ubiquitination by Deubiquitinases DupA and DupB
    Article Snippet: NEB Turbo (NEB, C2984H). .. T7 express cells were used for recombinant protein expression.

    Article Title: CRISPR-Cas9-Based Knockout of the Prion Protein and Its Effect on the Proteome
    Article Snippet: Paragraph title: Generation of gRNA Expression Vectors ... Oligonucleotides were phosphorylated with polynucleotide kinase (cat. no. EK0031; Fermentas, Ottawa, ON, Canada), annealed and inserted into the gRNA plasmid using T4 DNA ligase (cat no. M0202S; New England Biolabs) and transformed into Turbo competent E. coli (cat no. C2984H; New England Biolabs).

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells. .. All templates were cloned into a T7 expression plasmid system—the PURExpress control vector from New England Biolabs, called pNP1 in the text, pJL1, or pCOLADuet-1 (71406-3, Novagen)—using Gibson assembly ( ).

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells. .. All templates were cloned into a T7 expression plasmid system—the PURExpress control vector from New England Biolabs, called pNP1 in the text, pJL1, or pCOLADuet-1 (71406-3, Novagen)—using Gibson assembly ( ).

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: Transgenes were cloned into a pET21/28-derived bacterial expression vector. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Neq SSB-like protein Thermostable SSB from Nanoarchaeum equitans (Neq SSB-like protein) was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET29a(+) and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express Competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 30 °C, induced with 0.5 mM IPTG, expressed at 16 °C overnight (14 hr), and purified by IMAC FPLC using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences).

    Transformation Assay:

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo). .. Mutants were generated by QuikChange site-directed mutagenesis.

    Article Title: CRISPR-Cas9-Based Knockout of the Prion Protein and Its Effect on the Proteome
    Article Snippet: .. Oligonucleotides were phosphorylated with polynucleotide kinase (cat. no. EK0031; Fermentas, Ottawa, ON, Canada), annealed and inserted into the gRNA plasmid using T4 DNA ligase (cat no. M0202S; New England Biolabs) and transformed into Turbo competent E. coli (cat no. C2984H; New England Biolabs). .. Cell Culture and Transfection Mouse neuroblastoma Neuro-2a (N2a) (CCL-131) and mouse myoblast C2C12 cells (CRL-1772) were sourced from the American Tissue Culture Collection (ATCC) (Manassas, VA, USA).

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo). .. Mutants were generated by QuikChange site-directed mutagenesis.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Neq SSB-like protein Thermostable SSB from Nanoarchaeum equitans (Neq SSB-like protein) was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET29a(+) and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express Competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 30 °C, induced with 0.5 mM IPTG, expressed at 16 °C overnight (14 hr), and purified by IMAC FPLC using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences).

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction. .. Ampicillin resistant colonies were screened by PCR for the presence of the Atm cDNA fragment in 25 μl reactions (200 μM dNTP, 1.25 units JumpStart Taq DNA Polymerase [Sigma-Aldrich D9307], 0.5 μM of each primer).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Exons 2–10 of the Atm gene were then amplified from 1 μl of the RT-PCR reaction (∼500 ng starting RNA template) in 50 μl PCR reactions (250 μM dNTP, 1 μl PfuUltra II Fusion HS DNA Polymerase [Agilent Technologies 600670], 0.2 μM of each primer [5′-TGC TAG CAT GAG TCT AGC ACT CAA TGA TCT], [5′-TGC GGC CGC ACT AGA AGG TTT ACA]). .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Derivative Assay:

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: General template design and preparation DNA sequences encoding eforRed, dTomato, mOrange, ATF1, Ecarin, and Trx-Bx (batroxobin fused with thioredoxin as a solubility domain) genes were derived from the literature, codon-optimized for Escherichia coli , and synthesized as gBlocks or oligonucleotides by Integrated DNA Technologies. pPROEX-Aquamarine was a gift from F. Merola (plasmid #42889, Addgene), and pET29-sortaseA-penta-mutant was a gift from L. Griffith. .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells.

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: DNA sequences encoding eforRed, dTomato, mOrange, ATF1, Ecarin, and Trx-Bx (batroxobin fused with thioredoxin as a solubility domain) genes were derived from the literature, codon-optimized for Escherichia coli , and synthesized as gBlocks or oligonucleotides by Integrated DNA Technologies. pPROEX-Aquamarine was a gift from F. Merola (plasmid #42889, Addgene), and pET29-sortaseA-penta-mutant was a gift from L. Griffith. .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells.

    Flow Cytometry:

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Ligation:

    Article Title: CRISPR-Cas9-Based Knockout of the Prion Protein and Its Effect on the Proteome
    Article Snippet: The respective oligonucleotide pairs were obtained from Life Technologies (Burlington, ON, Canada) and were customized to include overhangs compatible for ligation into MLM3636 linearized by digestion with BsmB1 (cat. no. R0580S; New England BioLabs, Ipswich, MA, USA), a cloning site located in this vector on the 3′ side of a U6 promoter element. .. Oligonucleotides were phosphorylated with polynucleotide kinase (cat. no. EK0031; Fermentas, Ottawa, ON, Canada), annealed and inserted into the gRNA plasmid using T4 DNA ligase (cat no. M0202S; New England Biolabs) and transformed into Turbo competent E. coli (cat no. C2984H; New England Biolabs).

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction. .. Ampicillin resistant colonies were screened by PCR for the presence of the Atm cDNA fragment in 25 μl reactions (200 μM dNTP, 1.25 units JumpStart Taq DNA Polymerase [Sigma-Aldrich D9307], 0.5 μM of each primer).

    Solubility:

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: General template design and preparation DNA sequences encoding eforRed, dTomato, mOrange, ATF1, Ecarin, and Trx-Bx (batroxobin fused with thioredoxin as a solubility domain) genes were derived from the literature, codon-optimized for Escherichia coli , and synthesized as gBlocks or oligonucleotides by Integrated DNA Technologies. pPROEX-Aquamarine was a gift from F. Merola (plasmid #42889, Addgene), and pET29-sortaseA-penta-mutant was a gift from L. Griffith. .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells.

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: DNA sequences encoding eforRed, dTomato, mOrange, ATF1, Ecarin, and Trx-Bx (batroxobin fused with thioredoxin as a solubility domain) genes were derived from the literature, codon-optimized for Escherichia coli , and synthesized as gBlocks or oligonucleotides by Integrated DNA Technologies. pPROEX-Aquamarine was a gift from F. Merola (plasmid #42889, Addgene), and pET29-sortaseA-penta-mutant was a gift from L. Griffith. .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells.

    Generated:

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: C-terminal mCherry fusions with a (GGGS)2 linker were generated by Gibson cloning. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: C-terminal mCherry fusions with a (GGGS)2 linker were generated by Gibson cloning. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Polymerase Chain Reaction:

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: For protein expression, genes fragments encoding for BcLOV4 (GenBank accession number ), Cyphellophora europea LOV , Marsonnina brunnea LOV , Magnaporthe oryzae LOV , and Exophilia dermatitis LOV ( ) were ordered from Integrated DNA Technologies as gBlocks and assembled by Gibson cloning or PCR assembly. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: For protein expression, genes fragments encoding for BcLOV4 (GenBank accession number CCD53251.1), Cyphellophora europea LOV (ETN36999.1), Marsonnina brunnea LOV (EKD19672.1), Magnaporthe oryzae LOV (EHA46884.1), and Exophilia dermatitis LOV (EHY60539.1) were ordered from Integrated DNA Technologies as gBlocks and assembled by Gibson cloning or PCR assembly. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Exons 2–10 of the Atm gene were then amplified from 1 μl of the RT-PCR reaction (∼500 ng starting RNA template) in 50 μl PCR reactions (250 μM dNTP, 1 μl PfuUltra II Fusion HS DNA Polymerase [Agilent Technologies 600670], 0.2 μM of each primer [5′-TGC TAG CAT GAG TCT AGC ACT CAA TGA TCT], [5′-TGC GGC CGC ACT AGA AGG TTT ACA]). .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Sequencing:

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo). .. All sequences were verified by Sanger sequencing.

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo). .. All sequences were verified by Sanger sequencing.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Neq SSB-like protein Thermostable SSB from Nanoarchaeum equitans (Neq SSB-like protein) was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET29a(+) and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express Competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 30 °C, induced with 0.5 mM IPTG, expressed at 16 °C overnight (14 hr), and purified by IMAC FPLC using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences).

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Paragraph title: Atm cDNA sequencing ... NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Binding Assay:

    Article Title: Rational library design by functional CDR resampling
    Article Snippet: E. coli maltose binding protein (MBP) was produced in house. .. Turbo Competent E. coli (High Efficiency) cells (C2984I) and SOC media (B9020S) were purchased from New England Biolabs.

    Cellular Antioxidant Activity Assay:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Exons 2–10 of the Atm gene were then amplified from 1 μl of the RT-PCR reaction (∼500 ng starting RNA template) in 50 μl PCR reactions (250 μM dNTP, 1 μl PfuUltra II Fusion HS DNA Polymerase [Agilent Technologies 600670], 0.2 μM of each primer [5′-TGC TAG CAT GAG TCT AGC ACT CAA TGA TCT], [5′-TGC GGC CGC ACT AGA AGG TTT ACA]). .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Molecular Cloning:

    Article Title: Regulation of Phosphoribosyl-Linked Serine Ubiquitination by Deubiquitinases DupA and DupB
    Article Snippet: NEB Turbo (NEB, C2984H). .. NEB Turbo cells were used for plasmid amplification and molecular cloning.

    Fluorescence:

    Article Title: Plasmid-normalized quantification of relative mitochondrial DNA copy number
    Article Snippet: In order to verify whether the two fluorescent dyes present different intensities of fluorescence emission and to normalize for differences between runs (master mix performance, instrument calibration, environmental variability, etc.) a plasmid construct was designed. .. Plasmids were extracted from NEB Turbo Competent E. coli (New England Biolabs, Ipswich, MA, USA) with PureYield Plasmid Midiprep kit (Promega Co., Madison, WI, USA) and the concentration was measured using Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA).

    Mutagenesis:

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo). .. Mutants were generated by QuikChange site-directed mutagenesis.

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo). .. Mutants were generated by QuikChange site-directed mutagenesis.

    Isolation:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: RNA was isolated from the spleen of A-T [M] mice and converted to cDNA as described above. .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Purification:

    Article Title: Rational library design by functional CDR resampling
    Article Snippet: Biotinylated peptides antigens were designed in-house and purchased from Biopeptik with 90% purity (peptides: XIAP1B (Biotin-SGSPVSASTLARAGFLYTGE, Human), XIAP2B (Biotin-THADYLLRTGQVVDISDTIY, Human), GRAP2B (Biotin-SLNKLVDYYRTNSISRQKQI, Human), GRAP3B (Biotin-TDPVQLQAAGRVRWARALYD, Human), LMNA3B (Biotin-DEYQELLDIKLALDMEIHAYRK, Human), TDP_42_NLS (Biotin- PKD NKRKMDETDASSAVKVKRA), XIAP3B (Biotin-AEAVDKCPMCYTVITFKQK, Human), LMNA2B (Biotin-RIDSLSAQLSQLQKQLAAKE, Human) purified proteins were provided internally by Abcam (proteins: hIL-12p70( / , Human), FGF basic protein ( , Human), Dtk-Fc( , Human), BCMA-Fc(A7KBT3, Human), Nogo Fc( , Human), IL-13( , Human), myelin basic protein( , Human), hLeptin , IFN alpha2( , Human), BDNF( ,Human), IL-1 beta( , Human), IL3 beta( , Human). .. Turbo Competent E. coli (High Efficiency) cells (C2984I) and SOC media (B9020S) were purchased from New England Biolabs.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Neq SSB-like protein Thermostable SSB from Nanoarchaeum equitans (Neq SSB-like protein) was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET29a(+) and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express Competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 30 °C, induced with 0.5 mM IPTG, expressed at 16 °C overnight (14 hr), and purified by IMAC FPLC using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Exons 2–10 of the Atm gene were then amplified from 1 μl of the RT-PCR reaction (∼500 ng starting RNA template) in 50 μl PCR reactions (250 μM dNTP, 1 μl PfuUltra II Fusion HS DNA Polymerase [Agilent Technologies 600670], 0.2 μM of each primer [5′-TGC TAG CAT GAG TCT AGC ACT CAA TGA TCT], [5′-TGC GGC CGC ACT AGA AGG TTT ACA]). .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Protein Extraction:

    Article Title: Rational library design by functional CDR resampling
    Article Snippet: Turbo Competent E. coli (High Efficiency) cells (C2984I) and SOC media (B9020S) were purchased from New England Biolabs. .. B-Per Bacterial Protein Extraction Reagent (78248) was purchased from Pierce.

    Affinity Chromatography:

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Fast Protein Liquid Chromatography:

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: Expression and purification of Neq SSB-like protein Thermostable SSB from Nanoarchaeum equitans (Neq SSB-like protein) was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET29a(+) and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express Competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 30 °C, induced with 0.5 mM IPTG, expressed at 16 °C overnight (14 hr), and purified by IMAC FPLC using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences).

    CRISPR:

    Article Title: CRISPR-Cas9-Based Knockout of the Prion Protein and Its Effect on the Proteome
    Article Snippet: Suitable CRISPR target sites within Prnp Exon 3 positive and negative strands were identified using the ‘CRISPR Design Tool’ ( http://crispr.mit.edu/ ) described in the Results section. .. Oligonucleotides were phosphorylated with polynucleotide kinase (cat. no. EK0031; Fermentas, Ottawa, ON, Canada), annealed and inserted into the gRNA plasmid using T4 DNA ligase (cat no. M0202S; New England Biolabs) and transformed into Turbo competent E. coli (cat no. C2984H; New England Biolabs).

    Activated Clotting Time Assay:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Exons 2–10 of the Atm gene were then amplified from 1 μl of the RT-PCR reaction (∼500 ng starting RNA template) in 50 μl PCR reactions (250 μM dNTP, 1 μl PfuUltra II Fusion HS DNA Polymerase [Agilent Technologies 600670], 0.2 μM of each primer [5′-TGC TAG CAT GAG TCT AGC ACT CAA TGA TCT], [5′-TGC GGC CGC ACT AGA AGG TTT ACA]). .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Mouse Assay:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: RNA was isolated from the spleen of A-T [M] mice and converted to cDNA as described above. .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Plasmid Preparation:

    Article Title: Plasmid-normalized quantification of relative mitochondrial DNA copy number
    Article Snippet: .. Plasmids were extracted from NEB Turbo Competent E. coli (New England Biolabs, Ipswich, MA, USA) with PureYield Plasmid Midiprep kit (Promega Co., Madison, WI, USA) and the concentration was measured using Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). .. The plasmid was linearized by ScaI digestion (New England Biolabs, Ipswich, MA, USA) and the linearized plasmid was used as calibrator in all the qPCR experiments.

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: Transgenes were cloned into a pET21/28-derived bacterial expression vector. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: Regulation of Phosphoribosyl-Linked Serine Ubiquitination by Deubiquitinases DupA and DupB
    Article Snippet: NEB Turbo (NEB, C2984H). .. NEB Turbo cells were used for plasmid amplification and molecular cloning.

    Article Title: CRISPR-Cas9-Based Knockout of the Prion Protein and Its Effect on the Proteome
    Article Snippet: .. Oligonucleotides were phosphorylated with polynucleotide kinase (cat. no. EK0031; Fermentas, Ottawa, ON, Canada), annealed and inserted into the gRNA plasmid using T4 DNA ligase (cat no. M0202S; New England Biolabs) and transformed into Turbo competent E. coli (cat no. C2984H; New England Biolabs). .. Cell Culture and Transfection Mouse neuroblastoma Neuro-2a (N2a) (CCL-131) and mouse myoblast C2C12 cells (CRL-1772) were sourced from the American Tissue Culture Collection (ATCC) (Manassas, VA, USA).

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells. .. All templates were cloned into a T7 expression plasmid system—the PURExpress control vector from New England Biolabs, called pNP1 in the text, pJL1, or pCOLADuet-1 (71406-3, Novagen)—using Gibson assembly ( ).

    Article Title: BioBits™ Explorer: A modular synthetic biology education kit
    Article Snippet: .. Cloning and plasmid propagation were performed using either Mach1 (C862003, Thermo Fisher Scientific) or NEB Turbo (C2984H, New England Biolabs) competent E. coli cells. .. All templates were cloned into a T7 expression plasmid system—the PURExpress control vector from New England Biolabs, called pNP1 in the text, pJL1, or pCOLADuet-1 (71406-3, Novagen)—using Gibson assembly ( ).

    Article Title: Rpn (YhgA-Like) Proteins of Escherichia coli K-12 and Their Contribution to RecA-Independent Horizontal Transfer
    Article Snippet: Plasmids were prepared from E. coli Turbo cells (catalog number C2984; New England BioLabs [NEB]). .. For temperature-sensitive plasmids, incubation at 30°C was used to permit plasmid replication and incubation at 42°C was used to remove the plasmid.

    Article Title: Directly light-regulated binding of RGS-LOV photoreceptors to anionic membrane phospholipids
    Article Snippet: Transgenes were cloned into a pET21/28-derived bacterial expression vector. .. Genetic constructs were transformed into competent Escherichia coli (C2984H; NEB Turbo).

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: .. Expression and purification of Neq SSB-like protein Thermostable SSB from Nanoarchaeum equitans (Neq SSB-like protein) was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET29a(+) and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express Competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 30 °C, induced with 0.5 mM IPTG, expressed at 16 °C overnight (14 hr), and purified by IMAC FPLC using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences).

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Vector and insert were ligated using the Rapid DNA Ligation kit (Roche Applied Science 11635379001) as per the manufacturer’s recommended protocol. .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Electrophoresis:

    Article Title: A novel mouse model for ataxia-telangiectasia with a N-terminal mutation displays a behavioral defect and a low incidence of lymphoma but no increased oxidative burden
    Article Snippet: Amplified cDNA and vector were subjected to electrophoresis on 1% agarose gels and extracted using a QIAquick Gel Extraction kit (Qiagen 28706). .. NEB Turbo Competent E. coli (New England BioLabs C2984H) were transformed with 1 μl of ligation reaction.

    Functional Assay:

    Article Title: Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro
    Article Snippet: Expression and purification of Tt Ago Tt Ago was obtained as a synthetic, codon-optimized sequence with amino-terminal hexahistidine tag from GenScript in plasmid pET28c and transformed into NEB Turbo competent E. coli for cloning and propagation, and T7 Express lysY/Iq competent E. coli for expression. .. Large scale cultures were grown to OD600 0.6 at 37 °C, induced with 0.5 mM IPTG, expressed overnight (14 hr) at 16 °C, and purified by immobilized metal ion affinity chromatography (IMAC) using gravity flow with Nickel NTA Agarose Beads (Gold Biotechnology, St. Louis, Missouri), or fast protein liquid chromatography (FPLC) using a 5mL HisTrap FF column on an ÄKTA FPLC (GE Healthcare Life Sciences)—both methods were effective at producing functional, high-purity argonaute.

    Selection:

    Article Title: Engineering a Model Cell for Rational Tuning of GPCR Signaling
    Article Snippet: NEB® Turbo Competent E. coli was used for all cloning experiments. .. Selection and growth of E. coli was performed in Lysogeny Broth (LB) medium at 37°C with aeration.

    Produced:

    Article Title: Rational library design by functional CDR resampling
    Article Snippet: E. coli maltose binding protein (MBP) was produced in house. .. Turbo Competent E. coli (High Efficiency) cells (C2984I) and SOC media (B9020S) were purchased from New England Biolabs.

    Concentration Assay:

    Article Title: Plasmid-normalized quantification of relative mitochondrial DNA copy number
    Article Snippet: .. Plasmids were extracted from NEB Turbo Competent E. coli (New England Biolabs, Ipswich, MA, USA) with PureYield Plasmid Midiprep kit (Promega Co., Madison, WI, USA) and the concentration was measured using Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). .. The plasmid was linearized by ScaI digestion (New England Biolabs, Ipswich, MA, USA) and the linearized plasmid was used as calibrator in all the qPCR experiments.

    Recombinant:

    Article Title: Regulation of Phosphoribosyl-Linked Serine Ubiquitination by Deubiquitinases DupA and DupB
    Article Snippet: NEB Turbo (NEB, C2984H). .. T7 express cells were used for recombinant protein expression.

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