c3019 (New England Biolabs)

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New England Biolabs c3019
C3019, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/c3019/product/New England Biolabs
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99

c3019 - by Bioz Stars, 2021-04

86/100 stars

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Plasmid Preparation:

Article Title: Creation of Golden Gate constructs for gene doctoring
Article Snippet: We encourage further modification and sharing of these plasmids to broaden the available toolkit for Gene Doctoring of diverse targets and species with minimal effort. .. Bacterial strains E. coli NEB5α (New England BioLabs product № C2987) or NEB10β (New England BioLabs product № C3019) cells were used as hosts during plasmid construction. ..

Article Title: Targeted interplay between bacterial pathogens and host autophagy
Article Snippet: The resulting pET28-YhjJ plasmid was checked by sequencing. .. The resulting plasmid constructs were cloned into NEB® 10-beta Competent E. coli cells (New England Biolabs, C3019). .. Protein expression and GST affinity isolation pET28a-YhjJ, pGEX-4T1 (GE Healthcare Life Sciences, 28954549) and pGEX-4T1- MAP1LC3B/LC3B (gift from Dr Rob Layfield, School of Life Sciences, University of Nottingham) were transformed into Rosetta2(DE3) pLacI competent cells (Novagen, Merck Millipore 71404) that were grown on plates supplemented with kanamycin and chloramphenicol (pET28a-YhjJ) or ampicillin and chloramphenicol (pGEX-4T1 and pGEX-4T1-MAP1LC3B/LC3B).

Clone Assay:

Article Title: Single-cell measurement of plasmid copy number and promoter activity
Article Snippet: .. Strains and media E. coli strain NEB 10-beta [∆(ara-leu) 7697 araD139 fhuA ∆lacX74 galK16 galE15 e14- φ80dlacZ∆M15 recA1 relA1 endA1 nupG rpsL (Str R ) rph spoT1 ∆(mrr-hsdRMS-mcrBC) ] was used for all cloning and experiments (New England Biolabs, C3019). ..

Article Title: Control of nitrogen fixation in bacteria that associate with cereals.
Article Snippet: .. Legumes obtain nitrogen from air through rhizobia residing in root nodules. ..

Article Title: Harnessing the central dogma for stringent multi-level control of gene expression
Article Snippet: .. Strains, media and chemicals All cloning and characterization of genetic constructs was performed using Escherichia coli strain DH10-β (Δ(ara-leu) 7697 araD139 fhuA ΔlacX74 galK16 galE15 e14-ϕ80dlacZΔM15 recA1 relA1 endA1 nupG rpsL (StrR) rph spoT1 Δ(mrr-hsdRMS-mcrBC) (New England Biolabs, C3019I). .. Cells were grown in DH10-β outgrowth medium (New England Biolabs, B9035S) for transformation, LB broth (Sigma-Aldrich, L3522) for general propagation, and M9 minimal media supplemented with glucose (6.78 g/L Na2 HPO4 , 3 g/L KH2 PO4 , 1 g/L NH4 Cl, 0.5 g/L NaCl (Sigma-Aldrich, M6030), 0.34 g/L thiamine hydrochloride (Sigma T4625), 0.4% D-glucose (Sigma-Aldrich, G7528), 0.2% casamino acids (Acros, AC61204-5000), 2 mM MgSO4 (Acros, 213115000), and 0.1 mM CaCl2 (Sigma-Aldrich, C8106)) for characterization experiments.

Transformation Assay:

Article Title: Magnitude of stimulation dictates the cannabinoid-mediated differential T cell response to HIVgp120
Article Snippet: The IRES-neor cassette was amplified from plasmid pIRESneo3 (Clontech Laboratories, Mountain View, CA, USA) using primers IRES5′ and Neo3′ (Supplemental Table 1) with respective XhoI and HpaI sites. .. Each ligate was transformed into competent Escherichia coli cells, Stbl3 (Gibco Invitrogen) or 10-beta (C3019; New England Biolabs). ..

Construct:

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$As(V) detection involves an ArsC-independent reduction to As(III). (A) As(III) and As(V) detection by the pMP01 biosensor plasmid was compared for the E. coli host strain (NEB10-beta) and a deletion mutant lacking arsenate specific reductase (NEB10-beta Δ arsC ). Data points represent the mean output signal after 18 h of arsenic exposure in MGP of three independently grown cultures, quantified in triplicate. Error bars indicate standard deviations. We found no significant difference between wild-type and mutant strains ( p = 0.07). (B) Speciation analysis following addition of 32 μM As(V) to MGP exposure media (no cells), NEB10-beta cultures and NEB10-beta Δ arsC cultures. Treatments were subsampled, fractionated by centrifugation and analyzed before (left panel) and after (right panel) a 3-h incubation. As(III) and As(V) concentrations in pellet and supernatant fractions were quantified using HPLC-ICP-MS. The mean and standard deviation of three independent treatments are presented.$

Journal: Frontiers in Microbiology

Article Title: Insights Into Arsenite and Arsenate Uptake Pathways Using a Whole Cell Biosensor

doi: 10.3389/fmicb.2018.02310

Figure Lengend Snippet: As(V) detection involves an ArsC-independent reduction to As(III). (A) As(III) and As(V) detection by the pMP01 biosensor plasmid was compared for the E. coli host strain (NEB10-beta) and a deletion mutant lacking arsenate specific reductase (NEB10-beta Δ arsC ). Data points represent the mean output signal after 18 h of arsenic exposure in MGP of three independently grown cultures, quantified in triplicate. Error bars indicate standard deviations. We found no significant difference between wild-type and mutant strains ( p = 0.07). (B) Speciation analysis following addition of 32 μM As(V) to MGP exposure media (no cells), NEB10-beta cultures and NEB10-beta Δ arsC cultures. Treatments were subsampled, fractionated by centrifugation and analyzed before (left panel) and after (right panel) a 3-h incubation. As(III) and As(V) concentrations in pellet and supernatant fractions were quantified using HPLC-ICP-MS. The mean and standard deviation of three independent treatments are presented.

Article Snippet: The reporter plasmid was transformed into E. coli NEB10-beta (New England BioLabs) – a level 1, non-pathogenic, non-regulated host.

Techniques: Plasmid Preparation, Mutagenesis, Centrifugation, Incubation, High Performance Liquid Chromatography, Mass Spectrometry, Standard Deviation

Overview of the seven-part LCR of the toy-model plasmid by utilizing six bridging oligo-sets (BO-sets) with low crosstalk and six BO-sets with high crosstalk. Each BO-set was used five times to assemble the toy-model plasmid ( Figure 1B ) by using the baseline conditions with 8% v/v DMSO/0.45 M betaine and without both detergents. For all LCRs, 3 µl were transformed by electroporation in 30 µl NEB ® 10- β E. coli cells. For more detailed results of each BO-set refer to Supplementary Figures S4 and S5 . ( A ) The baseline conditions with DMSO and betaine resulted in low efficiencies and low amounts of colonies. No correlation between crosstalk and BO performance was found. ( B ) LCRs without DMSO and betaine resulted in more colonies and higher efficiencies in comparison to the baseline conditions. The sequences of all BO-sets are shown in Supplementary Table S3 . BO, bridging oligo; CFU, colony forming unit; DMSO, dimethyl sulfoxide.

Journal: Synthetic Biology

Article Title: Optimization of the experimental parameters of the ligase cycling reaction

doi: 10.1093/synbio/ysz020

Figure Lengend Snippet: Overview of the seven-part LCR of the toy-model plasmid by utilizing six bridging oligo-sets (BO-sets) with low crosstalk and six BO-sets with high crosstalk. Each BO-set was used five times to assemble the toy-model plasmid ( Figure 1B ) by using the baseline conditions with 8% v/v DMSO/0.45 M betaine and without both detergents. For all LCRs, 3 µl were transformed by electroporation in 30 µl NEB ® 10- β E. coli cells. For more detailed results of each BO-set refer to Supplementary Figures S4 and S5 . ( A ) The baseline conditions with DMSO and betaine resulted in low efficiencies and low amounts of colonies. No correlation between crosstalk and BO performance was found. ( B ) LCRs without DMSO and betaine resulted in more colonies and higher efficiencies in comparison to the baseline conditions. The sequences of all BO-sets are shown in Supplementary Table S3 . BO, bridging oligo; CFU, colony forming unit; DMSO, dimethyl sulfoxide.

Article Snippet: For both plasmids, 3 µl per LCR were transformed in 30 µl NEB® 10-β E. coli cells.

Techniques: Plasmid Preparation, Transformation Assay, Electroporation

Plasmid elimination and transformation experiments. (A) bla KPC -positive 2011 NIH Clinical Center outbreak K. pneumoniae isolate MALDI-TOF MS trace (the asterisk denotes the pKpQIL_p019 MS peak). (B) Plasmid cure eliminates the pKpQIL_p019 MS peak. (C) Transformation of the plasmid-cured strain with a pKpQIL-containing plasmid preparation restores the pKpQIL_p019 MS peak (asterisk). (D) A bla KPC -negative clinical K. pneumoniae isolate lacks the pKpQIL_p019 MS peak. (E) Transformation with a pKpQIL plasmid preparation yields a pKpQIL_p019 MS peak (asterisk), demonstrating that the ability to express pKpQIL_p019 is not unique to the 2011 NIH outbreak isolates. (F) A competent 10-beta E. coli strain lacks the pKpQIL_p019 MS peak. (G) Transformation with a pKpQIL plasmid preparation yields a pKpQIL_p019 MS peak (asterisk), demonstrating that the ability to express pKpQIL_p019 is not specific to the genus Klebsiella .

Journal: Journal of Clinical Microbiology

Article Title: A Rapid Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry-Based Method for Single-Plasmid Tracking in an Outbreak of Carbapenem-Resistant Enterobacteriaceae

doi: 10.1128/JCM.00694-14

Figure Lengend Snippet: Plasmid elimination and transformation experiments. (A) bla KPC -positive 2011 NIH Clinical Center outbreak K. pneumoniae isolate MALDI-TOF MS trace (the asterisk denotes the pKpQIL_p019 MS peak). (B) Plasmid cure eliminates the pKpQIL_p019 MS peak. (C) Transformation of the plasmid-cured strain with a pKpQIL-containing plasmid preparation restores the pKpQIL_p019 MS peak (asterisk). (D) A bla KPC -negative clinical K. pneumoniae isolate lacks the pKpQIL_p019 MS peak. (E) Transformation with a pKpQIL plasmid preparation yields a pKpQIL_p019 MS peak (asterisk), demonstrating that the ability to express pKpQIL_p019 is not unique to the 2011 NIH outbreak isolates. (F) A competent 10-beta E. coli strain lacks the pKpQIL_p019 MS peak. (G) Transformation with a pKpQIL plasmid preparation yields a pKpQIL_p019 MS peak (asterisk), demonstrating that the ability to express pKpQIL_p019 is not specific to the genus Klebsiella .

Article Snippet: A 10-beta chemically competent Escherichia coli strain (New England Biolabs [NEB], Ipswich, MA) ( ) was transformed with extracted plasmid DNA according to the manufacturer's instructions and selected on RambaChrom KPC agar.

Techniques: Plasmid Preparation, Transformation Assay, Mass Spectrometry