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Annexin V staining demonstrates the apoptotic effect of bacterially delivered violacein at a MOI of 25 at 10 h post-infection. ( a ) 7% of HeLa S3 cells treated with a control invasive <t>DH10B</t> showed annexin V staining (right panel); ( b ) 33% of HeLa S3 cells treated with bacterially delivered violacein showed annexin V staining. Scale bars represent 100 μm.
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Images

1) Product Images from "A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds"

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds

Journal: Micromachines

doi: 10.3390/mi8080230

Annexin V staining demonstrates the apoptotic effect of bacterially delivered violacein at a MOI of 25 at 10 h post-infection. ( a ) 7% of HeLa S3 cells treated with a control invasive DH10B showed annexin V staining (right panel); ( b ) 33% of HeLa S3 cells treated with bacterially delivered violacein showed annexin V staining. Scale bars represent 100 μm.
Figure Legend Snippet: Annexin V staining demonstrates the apoptotic effect of bacterially delivered violacein at a MOI of 25 at 10 h post-infection. ( a ) 7% of HeLa S3 cells treated with a control invasive DH10B showed annexin V staining (right panel); ( b ) 33% of HeLa S3 cells treated with bacterially delivered violacein showed annexin V staining. Scale bars represent 100 μm.

Techniques Used: Staining, Infection

2) Product Images from "Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome"

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome

Journal: Journal of Biomedicine and Biotechnology

doi: 10.1155/2011/926258

The genomic structures of the MHV-68 recombinant viruses and the MHV-68 BAC plasmid. (a) Schematic diagram of the construction of the MHV-68 BAC virus. Shown is the left end of the viral genome encompassing terminal repeats and ORFs M1–M3. The BAC vector sequence and a puromycin expression cassette was recombined into the locus where the EGFP expression cassette was inserted in MHV-68(EGFP). MHV-68(BAC) was selected for the loss of the EGFP expression. The pMHV-68(BAC) plasmid was obtained by electroporating the circular genome of MHV-68(BAC) into DH10B cells. Following transfection of pMHV-68(BAC) into BHK-21 cells, the reconstituted virus, pMHV-68(BAC)v, was generated. The structures of pMHV-68(BAC) and pMHV-68(BAC)v are identical to MHV-68(BAC), except that pMHV-68(BAC) is circular with termini fused. To remove the BAC insert flanking by loxP sites, pMHV-68(BAC)v was co-transfected with a Cre recombinase expression plasmid and MHV-68(loxP) was isolated. The sites of Hind III are indicated with H and the ones of EcoR I were indicated with E. (b) Restriction enzyme analysis of the genome of MHV-68 recombinant viruses and the MHV-68 BAC plasmid. Each DNA sample was digested with either EcoR I or Hind III. Lane 1 is virion DNA of wild-type MHV-68; lane 2 is virion DNA of MHV-68(BAC); lane 3 is the plasmid DNA of pMHV-68(BAC); lane 4 is virion DNA of pMHV-68(BAC)v; lane 5 is virion DNA of MHV-68(loxP) and lane 6 is virion DNA of MHV-68(EGFP). The wild-type fragments which were eliminated due to the insertion of the BAC sequence were marked with asterisks. The fragments marked with a, b, d, and e, were generated because of the BAC insert. The fragments marked with c and f, resulted from the removal of BAC, 200 bp larger than the fragments indicated with asterisks. The fragments marked with F were from the fused termini of the circular pMHV-68(BAC). On the bottom of each agarose gel picture is the corresponding Southern blot using the probe spanning nt 51–6298. The laddering patterns in EcoR I-digested samples were due to various number of repeats on the linear virion DNA but not present in pMHV-68(BAC) (lane 3).
Figure Legend Snippet: The genomic structures of the MHV-68 recombinant viruses and the MHV-68 BAC plasmid. (a) Schematic diagram of the construction of the MHV-68 BAC virus. Shown is the left end of the viral genome encompassing terminal repeats and ORFs M1–M3. The BAC vector sequence and a puromycin expression cassette was recombined into the locus where the EGFP expression cassette was inserted in MHV-68(EGFP). MHV-68(BAC) was selected for the loss of the EGFP expression. The pMHV-68(BAC) plasmid was obtained by electroporating the circular genome of MHV-68(BAC) into DH10B cells. Following transfection of pMHV-68(BAC) into BHK-21 cells, the reconstituted virus, pMHV-68(BAC)v, was generated. The structures of pMHV-68(BAC) and pMHV-68(BAC)v are identical to MHV-68(BAC), except that pMHV-68(BAC) is circular with termini fused. To remove the BAC insert flanking by loxP sites, pMHV-68(BAC)v was co-transfected with a Cre recombinase expression plasmid and MHV-68(loxP) was isolated. The sites of Hind III are indicated with H and the ones of EcoR I were indicated with E. (b) Restriction enzyme analysis of the genome of MHV-68 recombinant viruses and the MHV-68 BAC plasmid. Each DNA sample was digested with either EcoR I or Hind III. Lane 1 is virion DNA of wild-type MHV-68; lane 2 is virion DNA of MHV-68(BAC); lane 3 is the plasmid DNA of pMHV-68(BAC); lane 4 is virion DNA of pMHV-68(BAC)v; lane 5 is virion DNA of MHV-68(loxP) and lane 6 is virion DNA of MHV-68(EGFP). The wild-type fragments which were eliminated due to the insertion of the BAC sequence were marked with asterisks. The fragments marked with a, b, d, and e, were generated because of the BAC insert. The fragments marked with c and f, resulted from the removal of BAC, 200 bp larger than the fragments indicated with asterisks. The fragments marked with F were from the fused termini of the circular pMHV-68(BAC). On the bottom of each agarose gel picture is the corresponding Southern blot using the probe spanning nt 51–6298. The laddering patterns in EcoR I-digested samples were due to various number of repeats on the linear virion DNA but not present in pMHV-68(BAC) (lane 3).

Techniques Used: Recombinant, BAC Assay, Plasmid Preparation, Sequencing, Expressing, Transfection, Generated, Isolation, Agarose Gel Electrophoresis, Southern Blot

3) Product Images from "A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds"

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds

Journal: Micromachines

doi: 10.3390/mi8080230

Annexin V staining demonstrates the apoptotic effect of bacterially delivered violacein at a MOI of 25 at 10 h post-infection. ( a ) 7% of HeLa S3 cells treated with a control invasive DH10B showed annexin V staining (right panel); ( b ) 33% of HeLa S3 cells treated with bacterially delivered violacein showed annexin V staining. Scale bars represent 100 μm.
Figure Legend Snippet: Annexin V staining demonstrates the apoptotic effect of bacterially delivered violacein at a MOI of 25 at 10 h post-infection. ( a ) 7% of HeLa S3 cells treated with a control invasive DH10B showed annexin V staining (right panel); ( b ) 33% of HeLa S3 cells treated with bacterially delivered violacein showed annexin V staining. Scale bars represent 100 μm.

Techniques Used: Staining, Infection

4) Product Images from "Development of a Native Escherichia coli Induction System for Ionic Liquid Tolerance"

Article Title: Development of a Native Escherichia coli Induction System for Ionic Liquid Tolerance

Journal: PLoS ONE

doi: 10.1371/journal.pone.0101115

Growth of different eilA expression strains at increasing [C 2 mim]Cl concentrations. A–E: growth assays of E. coli DH10B carrying different promoter- eilA constructs. Due to day-to-day variability in the final cell density reached in the microtiter-plate experiments, growth curves were normalized to a start OD of 0 and a maximum OD of 0.5. A) pP ydfO’ - eilA , B) pP ydfA’-eilA , C) pP marR’-eilA , D) pP lacUV5-eilA, E) pP- eilA . Error bars represent standard errors. Blue: 0 mM [C 2 mim]Cl, red: 100 mM [C 2 mim]Cl, green: 200 mM [C 2 mim]Cl, purple: 400 mM [C 2 mim]Cl.
Figure Legend Snippet: Growth of different eilA expression strains at increasing [C 2 mim]Cl concentrations. A–E: growth assays of E. coli DH10B carrying different promoter- eilA constructs. Due to day-to-day variability in the final cell density reached in the microtiter-plate experiments, growth curves were normalized to a start OD of 0 and a maximum OD of 0.5. A) pP ydfO’ - eilA , B) pP ydfA’-eilA , C) pP marR’-eilA , D) pP lacUV5-eilA, E) pP- eilA . Error bars represent standard errors. Blue: 0 mM [C 2 mim]Cl, red: 100 mM [C 2 mim]Cl, green: 200 mM [C 2 mim]Cl, purple: 400 mM [C 2 mim]Cl.

Techniques Used: Expressing, Construct

5) Product Images from "The DNA strand of chimeric RNA/DNA oligonucleotides can direct gene repair/conversion activity in mammalian and plant cell-free extracts"

Article Title: The DNA strand of chimeric RNA/DNA oligonucleotides can direct gene repair/conversion activity in mammalian and plant cell-free extracts

Journal: Nucleic Acids Research

doi:

Genetic readout system for correction of a point mutation in plasmid pK s m4021. A mutant kanamycin gene harbored in plasmid pK s m4021 is the target for correction by oligonucleotides. The mutant G is converted to a C by the action of the oligo. Corrected plasmids confer resistance to kanamycin in E.coli strain DH10B after electroporation leading to the genetic readout and colony counts.
Figure Legend Snippet: Genetic readout system for correction of a point mutation in plasmid pK s m4021. A mutant kanamycin gene harbored in plasmid pK s m4021 is the target for correction by oligonucleotides. The mutant G is converted to a C by the action of the oligo. Corrected plasmids confer resistance to kanamycin in E.coli strain DH10B after electroporation leading to the genetic readout and colony counts.

Techniques Used: Mutagenesis, Plasmid Preparation, Electroporation

6) Product Images from "Biochemical Characterization of Pathogenic Mutations in Human Mitochondrial Methionyl-tRNA Formyltransferase *"

Article Title: Biochemical Characterization of Pathogenic Mutations in Human Mitochondrial Methionyl-tRNA Formyltransferase *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M114.610626

Analysis of the effect of mutations in MTF s in vivo . A and B , growth of E. coli DH10B co-transformants, containing pRSVCATam 1.2.5 and one of the pACD MTF s constructs, on 2× YT agar plates containing 50 μg/ml chloramphenicol. The growth
Figure Legend Snippet: Analysis of the effect of mutations in MTF s in vivo . A and B , growth of E. coli DH10B co-transformants, containing pRSVCATam 1.2.5 and one of the pACD MTF s constructs, on 2× YT agar plates containing 50 μg/ml chloramphenicol. The growth

Techniques Used: In Vivo, Construct

Related Articles

Clone Assay:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: .. The violacein operon (VioA-E) was cloned into the pUC19 vector (New England Biolabs, Ipswich, MA, USA) and used to transform DH10B (ThermoFisher Scientific, Waltham, MA, USA) cells previously transformed with plasmid TRIP 3.0 and maintained in growth media containing 50 μg/mL kanamycin and 100 μg/mL carbenicillin for dual expression of invasin and violacein. ..

Article Title: Biochemical Characterization of Pathogenic Mutations in Human Mitochondrial Methionyl-tRNA Formyltransferase *
Article Snippet: .. E. coli DH5α ( ) was used as host for cloning, and DH10B (Invitrogen) was used for in vivo assays. .. E. coli JM109 ( ) and BL21 (DE3) ( ) served as the host strains for overexpression of the recombinant forms of E. coli MTF and human mt-MTF variants, respectively.

Article Title: Cyclic GMP controls R. centenum cyst development
Article Snippet: .. E. coli strains DH5α, DH10B and SCS1101 were used for general cloning, BL21 Rosetta II (Invitrogen) for protein overexpression and S17-1 (λpir) for plasmid conjugation. .. E. coli strains were grown and cultured on agar-solidified or liquid Luria-Bertani (LB) media at 37 or 16°C with appropriate, antibiotics.

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: .. DH10B (Gibco-BRL) and ET12567 (pUZ8002) ( ) were used as Escherichia coli hosts for cloning and conjugation, respectively. .. Cosmid vector pHZ1358 ( ) and fosmid vector pCC2FOS (Epicentre Biotechnologies) were used for constructing genomic library.

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements
Article Snippet: For cloning of large DNA segments, restriction fragments were resolved on 0.5% agarose gels stained with SYBR gold (Invitrogen, Carlsbad, CA), detected with a Dark Reader Transilluminator (Clare Chemical Research, Denver, CO), and purified using a QIAEX II gel extraction kit (Qiagen, Hilden, GER). .. Ligations, performed with standard procedures, were electroporated into DH10B (Invitrogen) or SURE (Stratagene, La Jolla, CA).

Article Title: Nitric Oxide Signaling and Transcriptional Control of Denitrification Genes in Pseudomonas stutzeri
Article Snippet: The Escherichia coli strains used for propagation of plasmids were DH10B (Gibco-BRL) and JM110 ( ). .. Vectors used for cloning and sequencing were pBluescript II SK (Stratagene), pUCP22 , and pBSL15 , with the neomycinphosphotransferase II ( nptII ) gene conferring resistance to kanamycin.

Article Title: Diverse genetic error modes constrain large-scale bio-based production
Article Snippet: E . coli TOP10, similar to DH10B (Invitrogen): F− mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara , leu) 7697 galU galK rpsL (StrR) endA1 nupG E . coli XL1 (Agilent): recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacI q Z∆M15 Tn10 (Tet r )] E . coli MDS42 (Scarab Genomics): MG1655 genome reduced for all ISs, fhuACDB , endA and more . .. Strain kle1#1 was constructed by lambda-red recombineering-based deletion of chromosomal murI with a kanR deletion fragment (oligos, Supplementary Table ) in TOP10 cells harboring pSIM6 and pools of cloned RBS-variable pMevT-murI (Supplementary Table , construction described below).

Luciferase:

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used. .. For reporter assays, 10 ng of a firefly luciferase construct driven by the ISRE (interferon-stimulated response element) promoter and 1 ng of a Renilla luciferase construct driven by the housekeeping PGK (phosphoglycerate kinase) promoter were transfected into 293T cells seeded in 48-well plates together with 100 ng of a protein expression plasmid of a viral gene.

Binding Assay:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: The Listeriolysin O (LLO) gene and an additional ribosome binding site (RBS) were incorporated immediately downstream of invasin that confers invasive properties to bacteria (see main text). .. The violacein operon (VioA-E) was cloned into the pUC19 vector (New England Biolabs, Ipswich, MA, USA) and used to transform DH10B (ThermoFisher Scientific, Waltham, MA, USA) cells previously transformed with plasmid TRIP 3.0 and maintained in growth media containing 50 μg/mL kanamycin and 100 μg/mL carbenicillin for dual expression of invasin and violacein.

Stable Transfection:

Article Title: CRISPR/Cas9-induced transgene insertion and telomere-associated truncation of a single human chromosome for chromosome engineering in CHO and A9 cells
Article Snippet: The 1 kb artificial telomere was fragile in certain E . coli strains; therefore, the pBS-TEL/Dq HisD series was introduced by electroporation and amplified in DH10B (Thermo Fisher, Waltham, USA), an E . .. Coli strain that could stably maintain the 1 kb artificial telomere.

Synthesized:

Article Title: The DNA strand of chimeric RNA/DNA oligonucleotides can direct gene repair/conversion activity in mammalian and plant cell-free extracts
Article Snippet: Chimeric oligonucleotides and single-stranded oligonucleotides (including those with the indicated modifications) were synthesized using available phosphoramidites on controlled pore glass supports. .. The E.coli strain, DH10B, was obtained from Life Technologies (Gaithersburg, MD); DH10B cells contain a mutation in the RECA gene ( recA ).

Construct:

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used. .. For reporter assays, 10 ng of a firefly luciferase construct driven by the ISRE (interferon-stimulated response element) promoter and 1 ng of a Renilla luciferase construct driven by the housekeeping PGK (phosphoglycerate kinase) promoter were transfected into 293T cells seeded in 48-well plates together with 100 ng of a protein expression plasmid of a viral gene.

Article Title: CRISPR/Cas9-induced transgene insertion and telomere-associated truncation of a single human chromosome for chromosome engineering in CHO and A9 cells
Article Snippet: To construct pBS-TEL/Dq HisD Fcy; Fur, pBS-TEL/Dq HisD AN was digested with NotI and treated with rapid alkaline phosphatase, then ligated with an approximately 2.5kb fragment of pSelect-Zeo-Fcy; Fur, digested with EagI. .. The 1 kb artificial telomere was fragile in certain E . coli strains; therefore, the pBS-TEL/Dq HisD series was introduced by electroporation and amplified in DH10B (Thermo Fisher, Waltham, USA), an E .

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: DH10B (Gibco-BRL) and ET12567 (pUZ8002) ( ) were used as Escherichia coli hosts for cloning and conjugation, respectively. .. Two pHZ1358-based cosmid genomic libraries, NS1 and NS2, and one pCC2FOS-based fosmid genomic library, NS3, had been constructed for this work. pJTU1278 and pJTU1289 derived from pHZ1358 were used for gene inactivation. pMD18-T (Takara) and pBlueScript II SK(+) (Stratagene) were used for DNA sequencing.

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements
Article Snippet: Paragraph title: Plasmid constructs ... Ligations, performed with standard procedures, were electroporated into DH10B (Invitrogen) or SURE (Stratagene, La Jolla, CA).

Article Title: Diverse genetic error modes constrain large-scale bio-based production
Article Snippet: E . coli K-12 parental strains below were used to construct the strains analyzed (Table ) using the specified plasmids (Table ). .. E . coli TOP10, similar to DH10B (Invitrogen): F− mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara , leu) 7697 galU galK rpsL (StrR) endA1 nupG E . coli XL1 (Agilent): recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacI q Z∆M15 Tn10 (Tet r )] E . coli MDS42 (Scarab Genomics): MG1655 genome reduced for all ISs, fhuACDB , endA and more .

Incubation:

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: After 20 h of incubation at 4 °C, the ligation mixture was examined for its usefulness for bacterial transformation by PCR detection of a site-specific fragment insertion using a duplex primer pairs. .. The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions.

Amplification:

Article Title: CRISPR/Cas9-induced transgene insertion and telomere-associated truncation of a single human chromosome for chromosome engineering in CHO and A9 cells
Article Snippet: .. The 1 kb artificial telomere was fragile in certain E . coli strains; therefore, the pBS-TEL/Dq HisD series was introduced by electroporation and amplified in DH10B (Thermo Fisher, Waltham, USA), an E . .. Coli strain that could stably maintain the 1 kb artificial telomere.

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements
Article Snippet: GFP was amplified by PCR from pEGFP-N1 (Clontech, Mountain View, CA) with primers that added terminal SgfI and SmaI sites ( ) and cloned into pLL1 at those sites. .. Ligations, performed with standard procedures, were electroporated into DH10B (Invitrogen) or SURE (Stratagene, La Jolla, CA).

Article Title: Extrachromosomal Recombinant Adeno-Associated Virus Vector Genomes Are Primarily Responsible for Stable Liver Transduction In Vivo
Article Snippet: .. All plasmids were amplified in either DH10B (Gibco BRL, Gaithersburg, Md.) or Sure (Stratagene, Cedar Creek, Tex.) bacteria. .. Six- to eight-week-old female C57BL/6 mice were obtained from either Jackson Laboratory (Bar Harbor, Maine) or Taconic (Germantown, N.Y.).

Activity Assay:

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used. .. The normalized value was obtained by dividing the firefly luciferase reading with the Renilla luciferase activity of each sample.

Expressing:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: .. The violacein operon (VioA-E) was cloned into the pUC19 vector (New England Biolabs, Ipswich, MA, USA) and used to transform DH10B (ThermoFisher Scientific, Waltham, MA, USA) cells previously transformed with plasmid TRIP 3.0 and maintained in growth media containing 50 μg/mL kanamycin and 100 μg/mL carbenicillin for dual expression of invasin and violacein. ..

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: To remove the BAC sequence, a Cre recombinase expression plasmid was co-transfected with the viral BAC plasmid. .. For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used.

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: The 5′-Olig2-IRES-DsRed-3′ cassette was then excised from the pOlig2cDNA-IRES2-DsRed2 expression plasmid using the Nhe I and the Mfe I restriction enzymes (Invitrogen), blunt-ended with Klenow large fragment polymerase I (Roche) and then ligated to Bam HI/Sal I digested, blunt-ended and treated by alkaline phosphatase (Promega) lentiviral backbone plasmid pRRL.SIN.cPPT.PGK/GFP.WPRE. .. The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions.

Article Title: Extrachromosomal Recombinant Adeno-Associated Virus Vector Genomes Are Primarily Responsible for Stable Liver Transduction In Vivo
Article Snippet: Sleeping Beauty (SB)-based transposon plasmid pT-EF1α-hF.IX, carrying the EF1α enhancer-promoter-driven hF.IX expression cassette, and the pCMV-SB and pCMV-mSB transposase plasmids, encoding wild-type and nonfunctional mutant SB transposase genes, respectively, under the control of the human cytomegalovirus immediate-early gene enhancer-promoter, have been described elsewhere ( , ). .. All plasmids were amplified in either DH10B (Gibco BRL, Gaithersburg, Md.) or Sure (Stratagene, Cedar Creek, Tex.) bacteria.

Article Title: Diverse genetic error modes constrain large-scale bio-based production
Article Snippet: E . coli TOP10, similar to DH10B (Invitrogen): F− mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara , leu) 7697 galU galK rpsL (StrR) endA1 nupG E . coli XL1 (Agilent): recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacI q Z∆M15 Tn10 (Tet r )] E . coli MDS42 (Scarab Genomics): MG1655 genome reduced for all ISs, fhuACDB , endA and more . .. Recombineering followed standard procedures in which cells containing pSIM6 + pMevT-murI with variable RBS were grown in selectable 2xYT at 30 °C to mid-exponential phase for 3–5 h and induced at 42 °C for recombinase expression for 20 min.

Transformation Assay:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: .. The violacein operon (VioA-E) was cloned into the pUC19 vector (New England Biolabs, Ipswich, MA, USA) and used to transform DH10B (ThermoFisher Scientific, Waltham, MA, USA) cells previously transformed with plasmid TRIP 3.0 and maintained in growth media containing 50 μg/mL kanamycin and 100 μg/mL carbenicillin for dual expression of invasin and violacein. ..

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions. .. Aliquots of the transformation suspensions were plated on LB agar medium supplemented with Ap (100 μg/ml, Sigma).

Article Title: Diverse genetic error modes constrain large-scale bio-based production
Article Snippet: E . coli TOP10, similar to DH10B (Invitrogen): F− mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara , leu) 7697 galU galK rpsL (StrR) endA1 nupG E . coli XL1 (Agilent): recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacI q Z∆M15 Tn10 (Tet r )] E . coli MDS42 (Scarab Genomics): MG1655 genome reduced for all ISs, fhuACDB , endA and more . .. Standard chemical transformation or electroporation was used for gene introduction.

Over Expression:

Article Title: Biochemical Characterization of Pathogenic Mutations in Human Mitochondrial Methionyl-tRNA Formyltransferase *
Article Snippet: E. coli DH5α ( ) was used as host for cloning, and DH10B (Invitrogen) was used for in vivo assays. .. E. coli JM109 ( ) and BL21 (DE3) ( ) served as the host strains for overexpression of the recombinant forms of E. coli MTF and human mt-MTF variants, respectively.

Article Title: Cyclic GMP controls R. centenum cyst development
Article Snippet: .. E. coli strains DH5α, DH10B and SCS1101 were used for general cloning, BL21 Rosetta II (Invitrogen) for protein overexpression and S17-1 (λpir) for plasmid conjugation. .. E. coli strains were grown and cultured on agar-solidified or liquid Luria-Bertani (LB) media at 37 or 16°C with appropriate, antibiotics.

Derivative Assay:

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: DH10B (Gibco-BRL) and ET12567 (pUZ8002) ( ) were used as Escherichia coli hosts for cloning and conjugation, respectively. .. Two pHZ1358-based cosmid genomic libraries, NS1 and NS2, and one pCC2FOS-based fosmid genomic library, NS3, had been constructed for this work. pJTU1278 and pJTU1289 derived from pHZ1358 were used for gene inactivation. pMD18-T (Takara) and pBlueScript II SK(+) (Stratagene) were used for DNA sequencing.

Electroporation:

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: .. For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used. .. For reporter assays, 10 ng of a firefly luciferase construct driven by the ISRE (interferon-stimulated response element) promoter and 1 ng of a Renilla luciferase construct driven by the housekeeping PGK (phosphoglycerate kinase) promoter were transfected into 293T cells seeded in 48-well plates together with 100 ng of a protein expression plasmid of a viral gene.

Article Title: CRISPR/Cas9-induced transgene insertion and telomere-associated truncation of a single human chromosome for chromosome engineering in CHO and A9 cells
Article Snippet: .. The 1 kb artificial telomere was fragile in certain E . coli strains; therefore, the pBS-TEL/Dq HisD series was introduced by electroporation and amplified in DH10B (Thermo Fisher, Waltham, USA), an E . .. Coli strain that could stably maintain the 1 kb artificial telomere.

Article Title: Diverse genetic error modes constrain large-scale bio-based production
Article Snippet: E . coli TOP10, similar to DH10B (Invitrogen): F− mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara , leu) 7697 galU galK rpsL (StrR) endA1 nupG E . coli XL1 (Agilent): recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacI q Z∆M15 Tn10 (Tet r )] E . coli MDS42 (Scarab Genomics): MG1655 genome reduced for all ISs, fhuACDB , endA and more . .. Standard chemical transformation or electroporation was used for gene introduction.

Transfection:

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: Paragraph title: 2.5. DNA Transfection and Electroporation ... For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used.

Electroporation Bacterial Transformation:

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: After 20 h of incubation at 4 °C, the ligation mixture was examined for its usefulness for bacterial transformation by PCR detection of a site-specific fragment insertion using a duplex primer pairs. .. The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions.

Ligation:

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: .. The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions. .. Aliquots of the transformation suspensions were plated on LB agar medium supplemented with Ap (100 μg/ml, Sigma).

Cell Culture:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: The violacein operon (VioA-E) was cloned into the pUC19 vector (New England Biolabs, Ipswich, MA, USA) and used to transform DH10B (ThermoFisher Scientific, Waltham, MA, USA) cells previously transformed with plasmid TRIP 3.0 and maintained in growth media containing 50 μg/mL kanamycin and 100 μg/mL carbenicillin for dual expression of invasin and violacein. .. RFP was produced constitutively by maintaining 1 mM Isopropyl β-d -1-thiogalactopyranoside (IPTG, Sigma-Aldrich, St. Louis, MO, USA) in all media from the time of bacterial colony inoculation through mammalian cell invasion and cell culture.

Article Title: Cyclic GMP controls R. centenum cyst development
Article Snippet: E. coli strains DH5α, DH10B and SCS1101 were used for general cloning, BL21 Rosetta II (Invitrogen) for protein overexpression and S17-1 (λpir) for plasmid conjugation. .. E. coli strains were grown and cultured on agar-solidified or liquid Luria-Bertani (LB) media at 37 or 16°C with appropriate, antibiotics.

DNA Sequencing:

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: DH10B (Gibco-BRL) and ET12567 (pUZ8002) ( ) were used as Escherichia coli hosts for cloning and conjugation, respectively. .. Two pHZ1358-based cosmid genomic libraries, NS1 and NS2, and one pCC2FOS-based fosmid genomic library, NS3, had been constructed for this work. pJTU1278 and pJTU1289 derived from pHZ1358 were used for gene inactivation. pMD18-T (Takara) and pBlueScript II SK(+) (Stratagene) were used for DNA sequencing.

Isolation:

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: S. nanchangensis NS3226, the wild-type producer for meilingmycin, was used for meilingmycin isolation and generation of mutant strains. .. DH10B (Gibco-BRL) and ET12567 (pUZ8002) ( ) were used as Escherichia coli hosts for cloning and conjugation, respectively.

Sequencing:

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: To remove the BAC sequence, a Cre recombinase expression plasmid was co-transfected with the viral BAC plasmid. .. For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used.

Article Title: Nitric Oxide Signaling and Transcriptional Control of Denitrification Genes in Pseudomonas stutzeri
Article Snippet: The Escherichia coli strains used for propagation of plasmids were DH10B (Gibco-BRL) and JM110 ( ). .. Vectors used for cloning and sequencing were pBluescript II SK (Stratagene), pUCP22 , and pBSL15 , with the neomycinphosphotransferase II ( nptII ) gene conferring resistance to kanamycin.

Recombinant:

Article Title: Biochemical Characterization of Pathogenic Mutations in Human Mitochondrial Methionyl-tRNA Formyltransferase *
Article Snippet: E. coli DH5α ( ) was used as host for cloning, and DH10B (Invitrogen) was used for in vivo assays. .. E. coli JM109 ( ) and BL21 (DE3) ( ) served as the host strains for overexpression of the recombinant forms of E. coli MTF and human mt-MTF variants, respectively.

Molecular Cloning:

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: Paragraph title: Molecular cloning ... The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions.

In Vivo:

Article Title: Biochemical Characterization of Pathogenic Mutations in Human Mitochondrial Methionyl-tRNA Formyltransferase *
Article Snippet: .. E. coli DH5α ( ) was used as host for cloning, and DH10B (Invitrogen) was used for in vivo assays. .. E. coli JM109 ( ) and BL21 (DE3) ( ) served as the host strains for overexpression of the recombinant forms of E. coli MTF and human mt-MTF variants, respectively.

Methylation:

Article Title: Extrachromosomal Recombinant Adeno-Associated Virus Vector Genomes Are Primarily Responsible for Stable Liver Transduction In Vivo
Article Snippet: Thus, methylation status at the Xho I site serves as a marker for vector genome replication. .. All plasmids were amplified in either DH10B (Gibco BRL, Gaithersburg, Md.) or Sure (Stratagene, Cedar Creek, Tex.) bacteria.

Mutagenesis:

Article Title: The DNA strand of chimeric RNA/DNA oligonucleotides can direct gene repair/conversion activity in mammalian and plant cell-free extracts
Article Snippet: .. The E.coli strain, DH10B, was obtained from Life Technologies (Gaithersburg, MD); DH10B cells contain a mutation in the RECA gene ( recA ). ..

Article Title: Mutational Analysis of the Arabidopsis Nucleotide Binding Site-Leucine-Rich Repeat Resistance Gene RPS2
Article Snippet: The rps2-101C Δ rpm1 double mutant ( rps2-101C/rps2-101C; Δ rpm1/Δrpm1 ) is a Col-0 × Nd-0 hybrid ( ). .. Escherichia coli strains DH5α and DH10B (Life Technologies, Bethesda, MD) were used for plasmid construction procedures.

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: S. nanchangensis NS3226, the wild-type producer for meilingmycin, was used for meilingmycin isolation and generation of mutant strains. .. DH10B (Gibco-BRL) and ET12567 (pUZ8002) ( ) were used as Escherichia coli hosts for cloning and conjugation, respectively.

Article Title: Nitric Oxide Signaling and Transcriptional Control of Denitrification Genes in Pseudomonas stutzeri
Article Snippet: The generation of strains MK220 ( nirF ::Gmr ) , MK418 ( nosR ::Kmr ) , and MRD235 ( dnrD ::Kmr ) ( ) by insertional mutagenesis was described previously. .. The Escherichia coli strains used for propagation of plasmids were DH10B (Gibco-BRL) and JM110 ( ).

Article Title: Extrachromosomal Recombinant Adeno-Associated Virus Vector Genomes Are Primarily Responsible for Stable Liver Transduction In Vivo
Article Snippet: Sleeping Beauty (SB)-based transposon plasmid pT-EF1α-hF.IX, carrying the EF1α enhancer-promoter-driven hF.IX expression cassette, and the pCMV-SB and pCMV-mSB transposase plasmids, encoding wild-type and nonfunctional mutant SB transposase genes, respectively, under the control of the human cytomegalovirus immediate-early gene enhancer-promoter, have been described elsewhere ( , ). .. All plasmids were amplified in either DH10B (Gibco BRL, Gaithersburg, Md.) or Sure (Stratagene, Cedar Creek, Tex.) bacteria.

Conjugation Assay:

Article Title: Cyclic GMP controls R. centenum cyst development
Article Snippet: .. E. coli strains DH5α, DH10B and SCS1101 were used for general cloning, BL21 Rosetta II (Invitrogen) for protein overexpression and S17-1 (λpir) for plasmid conjugation. .. E. coli strains were grown and cultured on agar-solidified or liquid Luria-Bertani (LB) media at 37 or 16°C with appropriate, antibiotics.

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: .. DH10B (Gibco-BRL) and ET12567 (pUZ8002) ( ) were used as Escherichia coli hosts for cloning and conjugation, respectively. .. Cosmid vector pHZ1358 ( ) and fosmid vector pCC2FOS (Epicentre Biotechnologies) were used for constructing genomic library.

Labeling:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: The violacein operon (VioA-E) was cloned into the pUC19 vector (New England Biolabs, Ipswich, MA, USA) and used to transform DH10B (ThermoFisher Scientific, Waltham, MA, USA) cells previously transformed with plasmid TRIP 3.0 and maintained in growth media containing 50 μg/mL kanamycin and 100 μg/mL carbenicillin for dual expression of invasin and violacein. .. DH10B double positive for pUC19-VioA-E and TRIP 3.0 were transformed with spectinomycin resistant inducible vector pJExpress T5-lacO RFP for labeling cells with red fluorescent protein.

Purification:

Article Title: Biochemical Characterization of Pathogenic Mutations in Human Mitochondrial Methionyl-tRNA Formyltransferase *
Article Snippet: E. coli DH5α ( ) was used as host for cloning, and DH10B (Invitrogen) was used for in vivo assays. .. E. coli B105 ( ) was used for overexpression and purification of E. coli initiator tRNA (tRNA2 fMet ).

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements
Article Snippet: For cloning of large DNA segments, restriction fragments were resolved on 0.5% agarose gels stained with SYBR gold (Invitrogen, Carlsbad, CA), detected with a Dark Reader Transilluminator (Clare Chemical Research, Denver, CO), and purified using a QIAEX II gel extraction kit (Qiagen, Hilden, GER). .. Ligations, performed with standard procedures, were electroporated into DH10B (Invitrogen) or SURE (Stratagene, La Jolla, CA).

Polymerase Chain Reaction:

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: After 20 h of incubation at 4 °C, the ligation mixture was examined for its usefulness for bacterial transformation by PCR detection of a site-specific fragment insertion using a duplex primer pairs. .. The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions.

Article Title: CRISPR/Cas9-induced transgene insertion and telomere-associated truncation of a single human chromosome for chromosome engineering in CHO and A9 cells
Article Snippet: The Fcy; Fur gene was PCR amplified from pSelect-Zeo-Fcy; Fur (Invitrogen, San Diego, USA) and digested with SalI and XhoI. .. The 1 kb artificial telomere was fragile in certain E . coli strains; therefore, the pBS-TEL/Dq HisD series was introduced by electroporation and amplified in DH10B (Thermo Fisher, Waltham, USA), an E .

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements
Article Snippet: GFP was amplified by PCR from pEGFP-N1 (Clontech, Mountain View, CA) with primers that added terminal SgfI and SmaI sites ( ) and cloned into pLL1 at those sites. .. Ligations, performed with standard procedures, were electroporated into DH10B (Invitrogen) or SURE (Stratagene, La Jolla, CA).

FACS:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: .. We dosed a suspension of HeLa S3 cells with violacein producing invasive DH10B in droplets to confirm if a drug producing operon could be detected and at least partially enriched above background in one cycle of cell sorting. ..

Staining:

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements
Article Snippet: For cloning of large DNA segments, restriction fragments were resolved on 0.5% agarose gels stained with SYBR gold (Invitrogen, Carlsbad, CA), detected with a Dark Reader Transilluminator (Clare Chemical Research, Denver, CO), and purified using a QIAEX II gel extraction kit (Qiagen, Hilden, GER). .. Ligations, performed with standard procedures, were electroporated into DH10B (Invitrogen) or SURE (Stratagene, La Jolla, CA).

Plasmid Preparation:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: .. The violacein operon (VioA-E) was cloned into the pUC19 vector (New England Biolabs, Ipswich, MA, USA) and used to transform DH10B (ThermoFisher Scientific, Waltham, MA, USA) cells previously transformed with plasmid TRIP 3.0 and maintained in growth media containing 50 μg/mL kanamycin and 100 μg/mL carbenicillin for dual expression of invasin and violacein. ..

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: To remove the BAC sequence, a Cre recombinase expression plasmid was co-transfected with the viral BAC plasmid. .. For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used.

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: The 5′-Olig2-IRES-DsRed-3′ cassette was then excised from the pOlig2cDNA-IRES2-DsRed2 expression plasmid using the Nhe I and the Mfe I restriction enzymes (Invitrogen), blunt-ended with Klenow large fragment polymerase I (Roche) and then ligated to Bam HI/Sal I digested, blunt-ended and treated by alkaline phosphatase (Promega) lentiviral backbone plasmid pRRL.SIN.cPPT.PGK/GFP.WPRE. .. The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions.

Article Title: CRISPR/Cas9-induced transgene insertion and telomere-associated truncation of a single human chromosome for chromosome engineering in CHO and A9 cells
Article Snippet: Paragraph title: Plasmid construction ... The 1 kb artificial telomere was fragile in certain E . coli strains; therefore, the pBS-TEL/Dq HisD series was introduced by electroporation and amplified in DH10B (Thermo Fisher, Waltham, USA), an E .

Article Title: Mutational Analysis of the Arabidopsis Nucleotide Binding Site-Leucine-Rich Repeat Resistance Gene RPS2
Article Snippet: .. Escherichia coli strains DH5α and DH10B (Life Technologies, Bethesda, MD) were used for plasmid construction procedures. .. The uidA mutant ( E. coli strain PK803) was a gift of E. Signer ( ).

Article Title: Cyclic GMP controls R. centenum cyst development
Article Snippet: .. E. coli strains DH5α, DH10B and SCS1101 were used for general cloning, BL21 Rosetta II (Invitrogen) for protein overexpression and S17-1 (λpir) for plasmid conjugation. .. E. coli strains were grown and cultured on agar-solidified or liquid Luria-Bertani (LB) media at 37 or 16°C with appropriate, antibiotics.

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: DH10B (Gibco-BRL) and ET12567 (pUZ8002) ( ) were used as Escherichia coli hosts for cloning and conjugation, respectively. .. Cosmid vector pHZ1358 ( ) and fosmid vector pCC2FOS (Epicentre Biotechnologies) were used for constructing genomic library.

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements
Article Snippet: Paragraph title: Plasmid constructs ... Ligations, performed with standard procedures, were electroporated into DH10B (Invitrogen) or SURE (Stratagene, La Jolla, CA).

Article Title: Extrachromosomal Recombinant Adeno-Associated Virus Vector Genomes Are Primarily Responsible for Stable Liver Transduction In Vivo
Article Snippet: Sleeping Beauty (SB)-based transposon plasmid pT-EF1α-hF.IX, carrying the EF1α enhancer-promoter-driven hF.IX expression cassette, and the pCMV-SB and pCMV-mSB transposase plasmids, encoding wild-type and nonfunctional mutant SB transposase genes, respectively, under the control of the human cytomegalovirus immediate-early gene enhancer-promoter, have been described elsewhere ( , ). .. All plasmids were amplified in either DH10B (Gibco BRL, Gaithersburg, Md.) or Sure (Stratagene, Cedar Creek, Tex.) bacteria.

Selection:

Article Title: Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3
Article Snippet: The positive ligation sample-PCR was then used to transform DH5α, Top10, DH10B, Stbl2 and Stbl3 chemically competent cells (Invitrogen) according to the manufacturer’s instructions. .. Following overnight incubation at 30 or 37 °C, single colonies were screened by duplex PCR using insert-insert and insert-vector primer pairs for the selection of clones harbouring the plasmid with the desired orientation of the 5′-Olig2-IRES-DsRed-3′ insert.

Produced:

Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
Article Snippet: The violacein operon (VioA-E) was cloned into the pUC19 vector (New England Biolabs, Ipswich, MA, USA) and used to transform DH10B (ThermoFisher Scientific, Waltham, MA, USA) cells previously transformed with plasmid TRIP 3.0 and maintained in growth media containing 50 μg/mL kanamycin and 100 μg/mL carbenicillin for dual expression of invasin and violacein. .. RFP was produced constitutively by maintaining 1 mM Isopropyl β-d -1-thiogalactopyranoside (IPTG, Sigma-Aldrich, St. Louis, MO, USA) in all media from the time of bacterial colony inoculation through mammalian cell invasion and cell culture.

Article Title: Extrachromosomal Recombinant Adeno-Associated Virus Vector Genomes Are Primarily Responsible for Stable Liver Transduction In Vivo
Article Snippet: AAV-EF1α-F.IX.PMT was produced using 293PMT cells that constitutively express the bacterial PaeR7 methyltransferase ( ). .. All plasmids were amplified in either DH10B (Gibco BRL, Gaithersburg, Md.) or Sure (Stratagene, Cedar Creek, Tex.) bacteria.

BAC Assay:

Article Title: Construction and Characterization of an Infectious Murine Gammaherpesivrus-68 Bacterial Artificial Chromosome
Article Snippet: To remove the BAC sequence, a Cre recombinase expression plasmid was co-transfected with the viral BAC plasmid. .. For electroporation of DH10B (Invitrogen), a Gene Pulser II system (Bio-rad) with 0.1-cm cuvettes (Fisher) was used.

Marker:

Article Title: Extrachromosomal Recombinant Adeno-Associated Virus Vector Genomes Are Primarily Responsible for Stable Liver Transduction In Vivo
Article Snippet: Thus, methylation status at the Xho I site serves as a marker for vector genome replication. .. All plasmids were amplified in either DH10B (Gibco BRL, Gaithersburg, Md.) or Sure (Stratagene, Cedar Creek, Tex.) bacteria.

Gel Extraction:

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements
Article Snippet: For cloning of large DNA segments, restriction fragments were resolved on 0.5% agarose gels stained with SYBR gold (Invitrogen, Carlsbad, CA), detected with a Dark Reader Transilluminator (Clare Chemical Research, Denver, CO), and purified using a QIAEX II gel extraction kit (Qiagen, Hilden, GER). .. Ligations, performed with standard procedures, were electroporated into DH10B (Invitrogen) or SURE (Stratagene, La Jolla, CA).

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    This is a mate paired library control from E coli DH10B for use with the SOLiD series of sequencing instruments For Research Use Only Not for use in diagnostics procedures
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    99
    Thermo Fisher dh10b
    Annexin V staining demonstrates the apoptotic effect of bacterially delivered violacein at a MOI of 25 at 10 h post-infection. ( a ) 7% of HeLa S3 cells treated with a control invasive <t>DH10B</t> showed annexin V staining (right panel); ( b ) 33% of HeLa S3 cells treated with bacterially delivered violacein showed annexin V staining. Scale bars represent 100 μm.
    Dh10b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dh10b/product/Thermo Fisher
    Average 99 stars, based on 33 article reviews
    Price from $9.99 to $1999.99
    dh10b - by Bioz Stars, 2020-02
    99/100 stars
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    99
    Thermo Fisher e coli dh10b
    SBD-homologous domains might function to recognize PT-DNA. a Multiple sequence alignment of representative ScoMcrA-SBD homologs from Streptomyces coelicolor (#1), Streptomyces gancidicus (#2), Escherichia coli (#3), and Morganella morganii (#4). The sulfur-recognizing residues are highlighted in yellow. The consensus sequence is shown at the bottom. Right: Domain organizations of these ScoMcrA homologs. b The surface of ScoMcrA-SBD is colored according to conservation scores, which shows that its sulfur-recognizing cavity is highly conserved. c Purified ScoMcrA-SBD domain homologs from Streptomyces gancidicus (#2), Escherichia coli (#3), and Morganella morganii (#4) specifically recognized PT-DNA with the sulfur atom in the R p , but not in the S p , configuration. d Mutation of P482N in the Escherichia coli SBD homolog significantly diminished, while mutation of P607N in the Streptomyces gancidicus SBD homolog disrupted, the association with PT-DNA with the core sequence G PS AAC. e Heterologous expression of scomcrA homologs from S. gancidicus (#2), E. coli (#3), and M. morganii (#4) restricted transfer of the dnd gene cluster from Salmonella enterica , which contains the genes encoding the “writer” proteins of DNA phosphorothioation. Transformation frequencies of empty pBluescript vector (PT − ) and that harboring the dnd gene cluster (PT + ) into E. coli <t>DH10B</t> expressing various scomcrA homologs are shown
    E Coli Dh10b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 102 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli dh10b/product/Thermo Fisher
    Average 99 stars, based on 102 article reviews
    Price from $9.99 to $1999.99
    e coli dh10b - by Bioz Stars, 2020-02
    99/100 stars
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    Image Search Results


    Annexin V staining demonstrates the apoptotic effect of bacterially delivered violacein at a MOI of 25 at 10 h post-infection. ( a ) 7% of HeLa S3 cells treated with a control invasive DH10B showed annexin V staining (right panel); ( b ) 33% of HeLa S3 cells treated with bacterially delivered violacein showed annexin V staining. Scale bars represent 100 μm.

    Journal: Micromachines

    Article Title: A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds

    doi: 10.3390/mi8080230

    Figure Lengend Snippet: Annexin V staining demonstrates the apoptotic effect of bacterially delivered violacein at a MOI of 25 at 10 h post-infection. ( a ) 7% of HeLa S3 cells treated with a control invasive DH10B showed annexin V staining (right panel); ( b ) 33% of HeLa S3 cells treated with bacterially delivered violacein showed annexin V staining. Scale bars represent 100 μm.

    Article Snippet: We dosed a suspension of HeLa S3 cells with violacein producing invasive DH10B in droplets to confirm if a drug producing operon could be detected and at least partially enriched above background in one cycle of cell sorting.

    Techniques: Staining, Infection

    SBD-homologous domains might function to recognize PT-DNA. a Multiple sequence alignment of representative ScoMcrA-SBD homologs from Streptomyces coelicolor (#1), Streptomyces gancidicus (#2), Escherichia coli (#3), and Morganella morganii (#4). The sulfur-recognizing residues are highlighted in yellow. The consensus sequence is shown at the bottom. Right: Domain organizations of these ScoMcrA homologs. b The surface of ScoMcrA-SBD is colored according to conservation scores, which shows that its sulfur-recognizing cavity is highly conserved. c Purified ScoMcrA-SBD domain homologs from Streptomyces gancidicus (#2), Escherichia coli (#3), and Morganella morganii (#4) specifically recognized PT-DNA with the sulfur atom in the R p , but not in the S p , configuration. d Mutation of P482N in the Escherichia coli SBD homolog significantly diminished, while mutation of P607N in the Streptomyces gancidicus SBD homolog disrupted, the association with PT-DNA with the core sequence G PS AAC. e Heterologous expression of scomcrA homologs from S. gancidicus (#2), E. coli (#3), and M. morganii (#4) restricted transfer of the dnd gene cluster from Salmonella enterica , which contains the genes encoding the “writer” proteins of DNA phosphorothioation. Transformation frequencies of empty pBluescript vector (PT − ) and that harboring the dnd gene cluster (PT + ) into E. coli DH10B expressing various scomcrA homologs are shown

    Journal: Nature Communications

    Article Title: Structural basis for the recognition of sulfur in phosphorothioated DNA

    doi: 10.1038/s41467-018-07093-1

    Figure Lengend Snippet: SBD-homologous domains might function to recognize PT-DNA. a Multiple sequence alignment of representative ScoMcrA-SBD homologs from Streptomyces coelicolor (#1), Streptomyces gancidicus (#2), Escherichia coli (#3), and Morganella morganii (#4). The sulfur-recognizing residues are highlighted in yellow. The consensus sequence is shown at the bottom. Right: Domain organizations of these ScoMcrA homologs. b The surface of ScoMcrA-SBD is colored according to conservation scores, which shows that its sulfur-recognizing cavity is highly conserved. c Purified ScoMcrA-SBD domain homologs from Streptomyces gancidicus (#2), Escherichia coli (#3), and Morganella morganii (#4) specifically recognized PT-DNA with the sulfur atom in the R p , but not in the S p , configuration. d Mutation of P482N in the Escherichia coli SBD homolog significantly diminished, while mutation of P607N in the Streptomyces gancidicus SBD homolog disrupted, the association with PT-DNA with the core sequence G PS AAC. e Heterologous expression of scomcrA homologs from S. gancidicus (#2), E. coli (#3), and M. morganii (#4) restricted transfer of the dnd gene cluster from Salmonella enterica , which contains the genes encoding the “writer” proteins of DNA phosphorothioation. Transformation frequencies of empty pBluescript vector (PT − ) and that harboring the dnd gene cluster (PT + ) into E. coli DH10B expressing various scomcrA homologs are shown

    Article Snippet: The pACYCDuet™-1 vector and its derivatives carrying FL E. coli , M. morganii , and S. gancidicus ScoMcrA homologs with E. coli promoters (pJTU1673, pJTU1674, and pJTU1675) were introduced to E. coli DH10B, and competent cells of the resulting strains were prepared using the standard calcium chloride protocol.

    Techniques: Sequencing, Purification, Mutagenesis, Expressing, Transformation Assay, Plasmid Preparation