xbai  (New England Biolabs)


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    Name:
    XbaI
    Description:
    XbaI 15 000 units
    Catalog Number:
    r0145l
    Price:
    290
    Category:
    Restriction Enzymes
    Size:
    15 000 units
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    Structured Review

    New England Biolabs xbai
    XbaI
    XbaI 15 000 units
    https://www.bioz.com/result/xbai/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    xbai - by Bioz Stars, 2021-03
    86/100 stars

    Images

    1) Product Images from "Acinetobacter baumannii Gastrointestinal Colonization Is Facilitated by Secretory IgA Which Is Reductively Dissociated by Bacterial Thioredoxin A"

    Article Title: Acinetobacter baumannii Gastrointestinal Colonization Is Facilitated by Secretory IgA Which Is Reductively Dissociated by Bacterial Thioredoxin A

    Journal: mBio

    doi: 10.1128/mBio.01298-18

    Generation of a trxA deletion mutant. A schematic representation of the WT A. baumannii Ci79 and Δ trxA mutant genome surrounding the trxA locus with predicted fragment sizes detected by probe following either HindIII (white arrowhead) or XbaI (black arrowhead) digestion as well as probe target region (dotted box) (A). Southern blot (B) and Western blot (C) analyses of genomic and protein extracts, respectively, from WT Ci79, ΔtrxA , and ΔtrxA c A. baumannii were subsequently performed. PCR amplification of the trxA gene locus was performed using DNA from WT Ci79, ΔtrxA , and ΔtrxA c A. baumannii isolates. The PCR amplicons were subsequently digested with SalI and subjected to agarose gel electrophoresis (C). Total bacterial proteins were separated by electrophoresis and visualized in a Coomassie blue-stained polyacrylamide gel (D, left panel) or probed with anti-TrxA antibody to detect TrxA expression (D, right panel). Molecular marker sizes for DNA (bp; B and C) and protein (kDa; D) are provided.
    Figure Legend Snippet: Generation of a trxA deletion mutant. A schematic representation of the WT A. baumannii Ci79 and Δ trxA mutant genome surrounding the trxA locus with predicted fragment sizes detected by probe following either HindIII (white arrowhead) or XbaI (black arrowhead) digestion as well as probe target region (dotted box) (A). Southern blot (B) and Western blot (C) analyses of genomic and protein extracts, respectively, from WT Ci79, ΔtrxA , and ΔtrxA c A. baumannii were subsequently performed. PCR amplification of the trxA gene locus was performed using DNA from WT Ci79, ΔtrxA , and ΔtrxA c A. baumannii isolates. The PCR amplicons were subsequently digested with SalI and subjected to agarose gel electrophoresis (C). Total bacterial proteins were separated by electrophoresis and visualized in a Coomassie blue-stained polyacrylamide gel (D, left panel) or probed with anti-TrxA antibody to detect TrxA expression (D, right panel). Molecular marker sizes for DNA (bp; B and C) and protein (kDa; D) are provided.

    Techniques Used: Mutagenesis, Southern Blot, Western Blot, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Electrophoresis, Staining, Expressing, Marker

    2) Product Images from "Construction of a Bioluminescent Labelling Plasmid Vector for Bifidobacteria"

    Article Title: Construction of a Bioluminescent Labelling Plasmid Vector for Bifidobacteria

    Journal: Korean Journal for Food Science of Animal Resources

    doi: 10.5851/kosfa.2018.e17

    Restriction of pTG262 ( luc + ) (A) and physical genetic map of pTG262::pFI2576 rep ( luc + ) (B). M: HyperLadder™ 1 kb (Bioline Reagents Ltd.); 1: pTG262 [CCC (covalently closed circular) type]; 2: pTG262 ( luc + ) (CCC type); 3: pTG262 ( luc + ) (EcoRI-XbaI).
    Figure Legend Snippet: Restriction of pTG262 ( luc + ) (A) and physical genetic map of pTG262::pFI2576 rep ( luc + ) (B). M: HyperLadder™ 1 kb (Bioline Reagents Ltd.); 1: pTG262 [CCC (covalently closed circular) type]; 2: pTG262 ( luc + ) (CCC type); 3: pTG262 ( luc + ) (EcoRI-XbaI).

    Techniques Used: Countercurrent Chromatography

    Related Articles

    Agarose Gel Electrophoresis:

    Article Title: Antimicrobial Resistance, Virulence Profiles and Molecular Subtypes of Salmonella enterica Serovars Typhi and Paratyphi A Blood Isolates from Kolkata, India during 2009-2013
    Article Snippet: .. The DNA plugs were digested with 40 U of Xba I (New England Biolabs, MA) at 37°C for 18 h. The digested DNA was run on 1% pulsed field certified agarose gel (Bio-Rad, Hercules, Calif.) prepared in 0.5x TBE buffer (Sigma) using CHEF DRIII (Bio-Rad) apparatus with an initial switch time of 2.2 sec and a final switch time of 63.8 sec at 6 V/cm for 24 h. The gel was stained with ethidium bromide (1 µg/ml, Sigma), destained with deionized water and PFGE profiles were observed with the UV trans-illuminator using GelDoc (Bio-Rad). ..

    Article Title: Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene
    Article Snippet: A Southern blot analysis was performed to confirm the transgene. .. The DNA were digested using XbaI and DraI (New England Biolabs, Beverly, MA, USA), separated in 1% agarose gel, and then transferred to a Hybond-N+ membrane (Amersham Biosciences) by using capillary diffusion. .. The blots were probed with the 232-bp PCR fragment of DsRed (forward, 5′- CGACATCCCCGACTACATGA-3′ , and reverse, 5′- TCCTGGGGGTACAGCTTCTC-3′ ), which was [p32 ]dCTP-labeled by using the random primed labeling method.

    Article Title: Acinetobacter baumannii Gastrointestinal Colonization Is Facilitated by Secretory IgA Which Is Reductively Dissociated by Bacterial Thioredoxin A
    Article Snippet: Genetic manipulation was confirmed by Southern blot analysis. .. Genomic DNAs from Ci79, Δ trxA , and Δ trxA c strains were isolated using the GeneJET genomic DNA purification kit (Thermo Scientific, Rockford, IL), digested with XbaI or HindIII (New England BioLabs), and run on an 0.8% agarose gel at 16 V until completion. .. DNA was transferred to a nylon Biodyne B membrane (Thermo Scientific, Rockford, IL) and UV-cross-linked for 5 min. A DNA probe amplified from WT A. baumannii Ci79 genomic DNA using the DN_Fw/DN_Rv primer set was biotin labeled using the North2South biotin random prime labeling kit (Thermo Scientific, Rockport, IL).

    Article Title: Broad Meloidogyne Resistance in Potato Based on RNA Interference of Effector Gene 16D10
    Article Snippet: .. For each plant line, 15 µg DNA was digested with 50 U Xba I (New England Biolabs, Ipswich, MA) for 16 hr at 37 ° C, then separated on a 0.8% agarose gel at 70 V for 16 hr and transferred to a GeneScreen Plus nylon membrane (PerkinElmer, Boston, MA) in 10× saline sodium citrate (SSC) buffer (1.5 M NaCl, 0.15 M sodium citrate, pH 7). .. The membrane was UV cross-linked and hybridized for 16 hr at 42 ° C with a [α-32 P] dATP labeled 16D10i-2 probe ( ) in hybridization buffer (50% deionized formamide, 0.1 mg/ml salmon sperm DNA, 1% sodium dodecyl sulfate (SDS), 1 M NaCl, 10% dextran sulfate).

    Size-exclusion Chromatography:

    Article Title: Antimicrobial Resistance, Virulence Profiles and Molecular Subtypes of Salmonella enterica Serovars Typhi and Paratyphi A Blood Isolates from Kolkata, India during 2009-2013
    Article Snippet: .. The DNA plugs were digested with 40 U of Xba I (New England Biolabs, MA) at 37°C for 18 h. The digested DNA was run on 1% pulsed field certified agarose gel (Bio-Rad, Hercules, Calif.) prepared in 0.5x TBE buffer (Sigma) using CHEF DRIII (Bio-Rad) apparatus with an initial switch time of 2.2 sec and a final switch time of 63.8 sec at 6 V/cm for 24 h. The gel was stained with ethidium bromide (1 µg/ml, Sigma), destained with deionized water and PFGE profiles were observed with the UV trans-illuminator using GelDoc (Bio-Rad). ..

    Staining:

    Article Title: Antimicrobial Resistance, Virulence Profiles and Molecular Subtypes of Salmonella enterica Serovars Typhi and Paratyphi A Blood Isolates from Kolkata, India during 2009-2013
    Article Snippet: .. The DNA plugs were digested with 40 U of Xba I (New England Biolabs, MA) at 37°C for 18 h. The digested DNA was run on 1% pulsed field certified agarose gel (Bio-Rad, Hercules, Calif.) prepared in 0.5x TBE buffer (Sigma) using CHEF DRIII (Bio-Rad) apparatus with an initial switch time of 2.2 sec and a final switch time of 63.8 sec at 6 V/cm for 24 h. The gel was stained with ethidium bromide (1 µg/ml, Sigma), destained with deionized water and PFGE profiles were observed with the UV trans-illuminator using GelDoc (Bio-Rad). ..

    Diffusion-based Assay:

    Article Title: Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene
    Article Snippet: A Southern blot analysis was performed to confirm the transgene. .. The DNA were digested using XbaI and DraI (New England Biolabs, Beverly, MA, USA), separated in 1% agarose gel, and then transferred to a Hybond-N+ membrane (Amersham Biosciences) by using capillary diffusion. .. The blots were probed with the 232-bp PCR fragment of DsRed (forward, 5′- CGACATCCCCGACTACATGA-3′ , and reverse, 5′- TCCTGGGGGTACAGCTTCTC-3′ ), which was [p32 ]dCTP-labeled by using the random primed labeling method.

    Pulsed-Field Gel:

    Article Title: Evidence of Metabolic Switching and Implications for Food Safety from the Phenome(s) of Salmonella enterica Serovar Typhimurium DT104 Cultured at Selected Points across the Pork Production Food Chain
    Article Snippet: .. Pulsed-field gel electrophoresis (PFGE) was performed using the restriction endonucleases XbaI and AvrII (New England BioLabs) by following the CDC PulseNet protocol ( ). .. Salmonella enterica serovar Braenderup H9812 (ATCC BAA-664) was used as the reference strain for the molecular size standard.

    Isolation:

    Article Title: Acinetobacter baumannii Gastrointestinal Colonization Is Facilitated by Secretory IgA Which Is Reductively Dissociated by Bacterial Thioredoxin A
    Article Snippet: Genetic manipulation was confirmed by Southern blot analysis. .. Genomic DNAs from Ci79, Δ trxA , and Δ trxA c strains were isolated using the GeneJET genomic DNA purification kit (Thermo Scientific, Rockford, IL), digested with XbaI or HindIII (New England BioLabs), and run on an 0.8% agarose gel at 16 V until completion. .. DNA was transferred to a nylon Biodyne B membrane (Thermo Scientific, Rockford, IL) and UV-cross-linked for 5 min. A DNA probe amplified from WT A. baumannii Ci79 genomic DNA using the DN_Fw/DN_Rv primer set was biotin labeled using the North2South biotin random prime labeling kit (Thermo Scientific, Rockport, IL).

    DNA Purification:

    Article Title: Acinetobacter baumannii Gastrointestinal Colonization Is Facilitated by Secretory IgA Which Is Reductively Dissociated by Bacterial Thioredoxin A
    Article Snippet: Genetic manipulation was confirmed by Southern blot analysis. .. Genomic DNAs from Ci79, Δ trxA , and Δ trxA c strains were isolated using the GeneJET genomic DNA purification kit (Thermo Scientific, Rockford, IL), digested with XbaI or HindIII (New England BioLabs), and run on an 0.8% agarose gel at 16 V until completion. .. DNA was transferred to a nylon Biodyne B membrane (Thermo Scientific, Rockford, IL) and UV-cross-linked for 5 min. A DNA probe amplified from WT A. baumannii Ci79 genomic DNA using the DN_Fw/DN_Rv primer set was biotin labeled using the North2South biotin random prime labeling kit (Thermo Scientific, Rockport, IL).

    Polymerase Chain Reaction:

    Article Title: A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism
    Article Snippet: PCR was performed using Q5 Hot Start High-Fidelity DNA Polymerase (NEB) with the following cycling parameters: 30 s of initial incubation at 98°C, 35 cycles at 98°C for 10 s and at 72°C for 2 min, and a final elongation step of 2 min at 72°C. .. The PCR product was purified using QIAquick Gel Extraction Kit (Qiagen), digested with Hind III (NEB) and Xba I (NEB), and cloned into the pUC119 plasmid vector. .. Approximately 30 plasmid clones for each sorted sIgM (−) population were sequenced using universal forward, reverse, and Ig heavy chain primers 3 and 4.

    Purification:

    Article Title: A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism
    Article Snippet: PCR was performed using Q5 Hot Start High-Fidelity DNA Polymerase (NEB) with the following cycling parameters: 30 s of initial incubation at 98°C, 35 cycles at 98°C for 10 s and at 72°C for 2 min, and a final elongation step of 2 min at 72°C. .. The PCR product was purified using QIAquick Gel Extraction Kit (Qiagen), digested with Hind III (NEB) and Xba I (NEB), and cloned into the pUC119 plasmid vector. .. Approximately 30 plasmid clones for each sorted sIgM (−) population were sequenced using universal forward, reverse, and Ig heavy chain primers 3 and 4.

    Gel Extraction:

    Article Title: A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism
    Article Snippet: PCR was performed using Q5 Hot Start High-Fidelity DNA Polymerase (NEB) with the following cycling parameters: 30 s of initial incubation at 98°C, 35 cycles at 98°C for 10 s and at 72°C for 2 min, and a final elongation step of 2 min at 72°C. .. The PCR product was purified using QIAquick Gel Extraction Kit (Qiagen), digested with Hind III (NEB) and Xba I (NEB), and cloned into the pUC119 plasmid vector. .. Approximately 30 plasmid clones for each sorted sIgM (−) population were sequenced using universal forward, reverse, and Ig heavy chain primers 3 and 4.

    Clone Assay:

    Article Title: A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism
    Article Snippet: PCR was performed using Q5 Hot Start High-Fidelity DNA Polymerase (NEB) with the following cycling parameters: 30 s of initial incubation at 98°C, 35 cycles at 98°C for 10 s and at 72°C for 2 min, and a final elongation step of 2 min at 72°C. .. The PCR product was purified using QIAquick Gel Extraction Kit (Qiagen), digested with Hind III (NEB) and Xba I (NEB), and cloned into the pUC119 plasmid vector. .. Approximately 30 plasmid clones for each sorted sIgM (−) population were sequenced using universal forward, reverse, and Ig heavy chain primers 3 and 4.

    Plasmid Preparation:

    Article Title: A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism
    Article Snippet: PCR was performed using Q5 Hot Start High-Fidelity DNA Polymerase (NEB) with the following cycling parameters: 30 s of initial incubation at 98°C, 35 cycles at 98°C for 10 s and at 72°C for 2 min, and a final elongation step of 2 min at 72°C. .. The PCR product was purified using QIAquick Gel Extraction Kit (Qiagen), digested with Hind III (NEB) and Xba I (NEB), and cloned into the pUC119 plasmid vector. .. Approximately 30 plasmid clones for each sorted sIgM (−) population were sequenced using universal forward, reverse, and Ig heavy chain primers 3 and 4.

    Article Title: Construction of a Bioluminescent Labelling Plasmid Vector for Bifidobacteria
    Article Snippet: PCR product was digested with SalI restriction endonuclease (New England Biolabs, Hitchin, UK) and inserted into pUC19 cloning vector digested with the same restriction enzyme. .. A luciferase gene (luc+ ) was liberated with XbaI and EcoRI restriction endonucleases (New England Biolabs) from pLuc2 recombinant plasmid and inserted into pTG262 vector. .. The cloned pFI2576 rep was digested with SalI enzyme and subcloned into recombinant pTG262 (luc+ ) digested with the same enzyme.

    Luciferase:

    Article Title: Construction of a Bioluminescent Labelling Plasmid Vector for Bifidobacteria
    Article Snippet: PCR product was digested with SalI restriction endonuclease (New England Biolabs, Hitchin, UK) and inserted into pUC19 cloning vector digested with the same restriction enzyme. .. A luciferase gene (luc+ ) was liberated with XbaI and EcoRI restriction endonucleases (New England Biolabs) from pLuc2 recombinant plasmid and inserted into pTG262 vector. .. The cloned pFI2576 rep was digested with SalI enzyme and subcloned into recombinant pTG262 (luc+ ) digested with the same enzyme.

    Recombinant:

    Article Title: Construction of a Bioluminescent Labelling Plasmid Vector for Bifidobacteria
    Article Snippet: PCR product was digested with SalI restriction endonuclease (New England Biolabs, Hitchin, UK) and inserted into pUC19 cloning vector digested with the same restriction enzyme. .. A luciferase gene (luc+ ) was liberated with XbaI and EcoRI restriction endonucleases (New England Biolabs) from pLuc2 recombinant plasmid and inserted into pTG262 vector. .. The cloned pFI2576 rep was digested with SalI enzyme and subcloned into recombinant pTG262 (luc+ ) digested with the same enzyme.

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    New England Biolabs restriction endonucleases xbai
    (A) <t>PFGE</t> analysis of S . Typhimurium DT104 using <t>XbaI</t> and AvrII. Isolate groups were color identified for simplicity as follows, and the color codes were maintained in all of the assays: reference strain (white); group A, environmental (green); group B,
    Restriction Endonucleases Xbai, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction endonucleases xbai/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction endonucleases xbai - by Bioz Stars, 2021-03
    99/100 stars
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    (A) PFGE analysis of S . Typhimurium DT104 using XbaI and AvrII. Isolate groups were color identified for simplicity as follows, and the color codes were maintained in all of the assays: reference strain (white); group A, environmental (green); group B,

    Journal: Applied and Environmental Microbiology

    Article Title: Evidence of Metabolic Switching and Implications for Food Safety from the Phenome(s) of Salmonella enterica Serovar Typhimurium DT104 Cultured at Selected Points across the Pork Production Food Chain

    doi: 10.1128/AEM.01041-13

    Figure Lengend Snippet: (A) PFGE analysis of S . Typhimurium DT104 using XbaI and AvrII. Isolate groups were color identified for simplicity as follows, and the color codes were maintained in all of the assays: reference strain (white); group A, environmental (green); group B,

    Article Snippet: Pulsed-field gel electrophoresis (PFGE) was performed using the restriction endonucleases XbaI and AvrII (New England BioLabs) by following the CDC PulseNet protocol ( ).

    Techniques:

    Mapping and tandem arrangement of DXZ4 in primates . (a) Direct-labeled fluorescence in situ hybridization (FISH) of human DXZ4 BAC clone (2272M5; red) and human PLS3 BAC clone (268A15; green) to male and female rhesus macaque metaphase chromosomes. White arrows point to the hybridizing X chromosome. Metaphase chromosomes were counterstained with DAPI, and converted to gray-scale to assist in visualizing the FISH signals. (b) Southern blot of Xba I digested primate genomic DNA separated by pulsed field gel electrophoresis, hybridized with a human digoxigenin-labeled DXZ4 probe. Primates and group are listed along the top and gender indicated by M (male) or F (female), including rhesus macaque (R. Macaque), pig-tailed macaque (P-T. Macaque), common squirrel monkey (Sq. Monkey) and black-handed spider monkey (Sp. Monkey). Size in kilobases is given to the left. (c) Ethidium bromide stained 0.9% agarose gel showing green monkey and macaque BAC DNA digested with the restriction endonuclease Hin dIII and separated by gel electrophoresis. The sizes of the molecular weight marker are given to the left in kilobases.

    Journal: Genome Biology

    Article Title: Characterization of DXZ4 conservation in primates implies important functional roles for CTCF binding, array expression and tandem repeat organization on the X chromosome

    doi: 10.1186/gb-2011-12-4-r37

    Figure Lengend Snippet: Mapping and tandem arrangement of DXZ4 in primates . (a) Direct-labeled fluorescence in situ hybridization (FISH) of human DXZ4 BAC clone (2272M5; red) and human PLS3 BAC clone (268A15; green) to male and female rhesus macaque metaphase chromosomes. White arrows point to the hybridizing X chromosome. Metaphase chromosomes were counterstained with DAPI, and converted to gray-scale to assist in visualizing the FISH signals. (b) Southern blot of Xba I digested primate genomic DNA separated by pulsed field gel electrophoresis, hybridized with a human digoxigenin-labeled DXZ4 probe. Primates and group are listed along the top and gender indicated by M (male) or F (female), including rhesus macaque (R. Macaque), pig-tailed macaque (P-T. Macaque), common squirrel monkey (Sq. Monkey) and black-handed spider monkey (Sp. Monkey). Size in kilobases is given to the left. (c) Ethidium bromide stained 0.9% agarose gel showing green monkey and macaque BAC DNA digested with the restriction endonuclease Hin dIII and separated by gel electrophoresis. The sizes of the molecular weight marker are given to the left in kilobases.

    Article Snippet: Agarose embedded DNA was digested with Xba I (NEB, Ipswich, MA, USA).

    Techniques: Labeling, Fluorescence, In Situ Hybridization, Fluorescence In Situ Hybridization, BAC Assay, Southern Blot, Pulsed-Field Gel, Electrophoresis, Staining, Agarose Gel Electrophoresis, Nucleic Acid Electrophoresis, Molecular Weight, Marker

    gRNA library construction using a semi-random primer. ( A ) Semi-random primer. Poly(A), polyadenylate. ( B ) Type III and type IIS restriction sites to cut out the 20-bp guide sequence. Ec, Eco P15I; Ac, Acu I. ( C ) Scheme of gRNA library construction. Bg, Bgl II; Xb, Xba I; Bs, Bsm BI; Aa, Aat II. PCR, polymerase chain reaction; lentiCRISPR v2, lentiCRISPR version 2. ( D ) Short-range PCR for PCR cycle optimization and size fractionation of the guide sequence. PCR products were run on 20% polyacrylamide gels. A 10-bp ladder was used as the size marker. Bands of the expected sizes are marked by triangles.

    Journal: Science Advances

    Article Title: A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism

    doi: 10.1126/sciadv.1600699

    Figure Lengend Snippet: gRNA library construction using a semi-random primer. ( A ) Semi-random primer. Poly(A), polyadenylate. ( B ) Type III and type IIS restriction sites to cut out the 20-bp guide sequence. Ec, Eco P15I; Ac, Acu I. ( C ) Scheme of gRNA library construction. Bg, Bgl II; Xb, Xba I; Bs, Bsm BI; Aa, Aat II. PCR, polymerase chain reaction; lentiCRISPR v2, lentiCRISPR version 2. ( D ) Short-range PCR for PCR cycle optimization and size fractionation of the guide sequence. PCR products were run on 20% polyacrylamide gels. A 10-bp ladder was used as the size marker. Bands of the expected sizes are marked by triangles.

    Article Snippet: The PCR product was purified using QIAquick Gel Extraction Kit (Qiagen), digested with Hind III (NEB) and Xba I (NEB), and cloned into the pUC119 plasmid vector.

    Techniques: Sequencing, Polymerase Chain Reaction, Fractionation, Marker

    Generation of a trxA deletion mutant. A schematic representation of the WT A. baumannii Ci79 and Δ trxA mutant genome surrounding the trxA locus with predicted fragment sizes detected by probe following either HindIII (white arrowhead) or XbaI (black arrowhead) digestion as well as probe target region (dotted box) (A). Southern blot (B) and Western blot (C) analyses of genomic and protein extracts, respectively, from WT Ci79, ΔtrxA , and ΔtrxA c A. baumannii were subsequently performed. PCR amplification of the trxA gene locus was performed using DNA from WT Ci79, ΔtrxA , and ΔtrxA c A. baumannii isolates. The PCR amplicons were subsequently digested with SalI and subjected to agarose gel electrophoresis (C). Total bacterial proteins were separated by electrophoresis and visualized in a Coomassie blue-stained polyacrylamide gel (D, left panel) or probed with anti-TrxA antibody to detect TrxA expression (D, right panel). Molecular marker sizes for DNA (bp; B and C) and protein (kDa; D) are provided.

    Journal: mBio

    Article Title: Acinetobacter baumannii Gastrointestinal Colonization Is Facilitated by Secretory IgA Which Is Reductively Dissociated by Bacterial Thioredoxin A

    doi: 10.1128/mBio.01298-18

    Figure Lengend Snippet: Generation of a trxA deletion mutant. A schematic representation of the WT A. baumannii Ci79 and Δ trxA mutant genome surrounding the trxA locus with predicted fragment sizes detected by probe following either HindIII (white arrowhead) or XbaI (black arrowhead) digestion as well as probe target region (dotted box) (A). Southern blot (B) and Western blot (C) analyses of genomic and protein extracts, respectively, from WT Ci79, ΔtrxA , and ΔtrxA c A. baumannii were subsequently performed. PCR amplification of the trxA gene locus was performed using DNA from WT Ci79, ΔtrxA , and ΔtrxA c A. baumannii isolates. The PCR amplicons were subsequently digested with SalI and subjected to agarose gel electrophoresis (C). Total bacterial proteins were separated by electrophoresis and visualized in a Coomassie blue-stained polyacrylamide gel (D, left panel) or probed with anti-TrxA antibody to detect TrxA expression (D, right panel). Molecular marker sizes for DNA (bp; B and C) and protein (kDa; D) are provided.

    Article Snippet: Genomic DNAs from Ci79, Δ trxA , and Δ trxA c strains were isolated using the GeneJET genomic DNA purification kit (Thermo Scientific, Rockford, IL), digested with XbaI or HindIII (New England BioLabs), and run on an 0.8% agarose gel at 16 V until completion.

    Techniques: Mutagenesis, Southern Blot, Western Blot, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Electrophoresis, Staining, Expressing, Marker