xbai  (Millipore)

 
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    • 95
    Name:
    Xba I
    Description:
    Xba I recognizes the sequence T↓ CTAGA and generates fragments with 5 cohesive termini The enzyme needs at least two nucleotides around the target sequence before it will cut the cleavage site Compatible endsXba I ends are compatible with fragments generated by Avr I Nhe I and Spe I IsoschizomersThe enzyme is not known to have isoschizomers Methylation sensitivityXba I digestion of DNA is inhibited by the dam gene product of E coli which methylates the 6N position of adenine in the sequence GATC The enzyme is also inhibited by 5 methylcytosine or 5 hydroxymethylcytosine at the site indicated on the recognition sequence Activity in SuRE Cut Buffer SystemBuffer printed in bold face type is the buffer recommended for optimal activity A B L M H100 75 100 75 100 75 100 100 Relative activity in complete PCR mixRelative activity in PCR mix Taq DNA Polymerase buffer is 60 The PCR mix contained DNA primers 10 mM Tris HCl pH 8 3 20C 50 mM KCl 1 5 mM MgCl2 200 M dNTPs 2 5 U Taq DNA polymerase The mix was subjected to 25 amplification cycles Activity in reaction buffer of Pwo SuperYield DNA Polymerase PCR Mix is 25 When supplemented with GC RICH Solution activity increases to 100 Incubation temperature 37CNumber of cleavage sites on different DNAslambda Ad2 SV40 X174 M13mp7 M13mp18 pBR322 pBR328 pUC181 5 0 0 0 1 0 0 1PFGE testedXba I has been tested in Pulsed Field Gel Electrophoresis on bacterial chromosomes For cleavage of genomic DNA E coli C 600 embedded in agarose for PFGE analysis we recommend 10 U enzyme per g DNA and approximately 4 hour incubation time Ligation and recutting assayXba I fragments obtained by complete digestion of 1 g pUC18 DNA are ligated with 1 U T4 DNA Ligase in a volume of 10 l by incubation for 16 hours at 4C in 66 mM Tris HCl 5 mM MgCl2 5 mM Dithiothreitol 1 mM ATP pH 7 5 at 20C resulting in 90 recovery of pUC18 DNA Subsequent re cutting with Xba I yields 95 of the typical pattern of pUC18 x Xba I fragments
    Catalog Number:
    xbai-ro
    Price:
    None
    Applications:
    The restriction enzyme Xba I has been used for the digestion of genomic DNA.
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    Structured Review

    Millipore xbai
    Xba I recognizes the sequence T↓ CTAGA and generates fragments with 5 cohesive termini The enzyme needs at least two nucleotides around the target sequence before it will cut the cleavage site Compatible endsXba I ends are compatible with fragments generated by Avr I Nhe I and Spe I IsoschizomersThe enzyme is not known to have isoschizomers Methylation sensitivityXba I digestion of DNA is inhibited by the dam gene product of E coli which methylates the 6N position of adenine in the sequence GATC The enzyme is also inhibited by 5 methylcytosine or 5 hydroxymethylcytosine at the site indicated on the recognition sequence Activity in SuRE Cut Buffer SystemBuffer printed in bold face type is the buffer recommended for optimal activity A B L M H100 75 100 75 100 75 100 100 Relative activity in complete PCR mixRelative activity in PCR mix Taq DNA Polymerase buffer is 60 The PCR mix contained DNA primers 10 mM Tris HCl pH 8 3 20C 50 mM KCl 1 5 mM MgCl2 200 M dNTPs 2 5 U Taq DNA polymerase The mix was subjected to 25 amplification cycles Activity in reaction buffer of Pwo SuperYield DNA Polymerase PCR Mix is 25 When supplemented with GC RICH Solution activity increases to 100 Incubation temperature 37CNumber of cleavage sites on different DNAslambda Ad2 SV40 X174 M13mp7 M13mp18 pBR322 pBR328 pUC181 5 0 0 0 1 0 0 1PFGE testedXba I has been tested in Pulsed Field Gel Electrophoresis on bacterial chromosomes For cleavage of genomic DNA E coli C 600 embedded in agarose for PFGE analysis we recommend 10 U enzyme per g DNA and approximately 4 hour incubation time Ligation and recutting assayXba I fragments obtained by complete digestion of 1 g pUC18 DNA are ligated with 1 U T4 DNA Ligase in a volume of 10 l by incubation for 16 hours at 4C in 66 mM Tris HCl 5 mM MgCl2 5 mM Dithiothreitol 1 mM ATP pH 7 5 at 20C resulting in 90 recovery of pUC18 DNA Subsequent re cutting with Xba I yields 95 of the typical pattern of pUC18 x Xba I fragments
    https://www.bioz.com/result/xbai/product/Millipore
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    xbai - by Bioz Stars, 2021-03
    95/100 stars

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    Related Articles

    Software:

    Article Title: Antimicrobial resistance and genetic diversity of Escherichiacoli isolated from humans and foods
    Article Snippet: .. Data Analysis The Xba I (Sigma-Aldrich, USA) fingerprints were analyzed using GelCompar II software (Applied Maths, Kortrijk, Belgium). .. A similarity dendrogram was constructed by the unweighted pair group (UPGMA) method using the Dice similarity coefficient; the relatedness between the isolates was interpreted according to the method described by .

    Clone Assay:

    Article Title: RNA Polymerase Activity and Specific RNA Structure Are Required for Efficient HCV Replication in Cultured Cells
    Article Snippet: Primer sequences for mutagenesis are listed in . .. Following digestion with Xba I and Xho I, DNA fragments were cloned into the Nhe I and Xho I sites of pET21b (Novagen, Madison, WI), resulting in pET21bHCVJFH-1RdRpwt and pET21bHCVJ6CFRdRpwt. pET21bHCVJFH-1RdRpwt and pET21bHCVJ6CFRdRpwt were then digested with Xba I and Xho I and the RdRP fragments cloned into the same restriction sites of pET28a, resulting in pET21(KM)JFH-1RdRpwt and pET21(KM)J6CFRdRpwt, respectively. .. Mutation analysis of J6CF and JFH-1 RdRP JFH-1-type substitutions (S377R, A450S, S455N, R517K, and Y561F in the NS5B region; amino acid numbers are based on the AA relative numbering ) were introduced into J6CF RdRP and J6CF-like substitutions (S450A, N455S, K517R, F561Y, and F561I) and D318A were introduced into JFH-1 RdRP using the QuickChange II Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA).

    Article Title: Human serum albumin-mediated apoptin delivery suppresses breast cancer cell growth in vitro and in vivo
    Article Snippet: The PCR fragments were cloned into pEGFP-N1 using the T4 DNA ligase (Sigma-Aldrich; Merck Millipore). .. Subsequently, the insert site was released from pEGFP-N1 by digestion with Eco RI and Xba I restriction enzymes (Sigma-Aldrich; Merck Millipore), and PCR fragments were cloned into pEGFP-N1 using the T4 DNA ligase; the complex was termed pEGFP-N1-Apoptin, which was amplified in bacterial cells (E. coli DH5α; Takara Bio, Inc., Otsu, Japan). .. The cells were transfected with pEGFP-N1-Apoptin as follows: Following thawing of competent cells, 2 µl pEGFP-N1-Apoptin was added to 100 µl competent cells and incubated on ice for 30 min.

    Amplification:

    Article Title: Oral Immunogenicity of Plant-Made Mycobacterium tuberculosis ESAT6 and CFP10
    Article Snippet: Amplification of esat6 and cfp10 Gene Sequences and Their Cloning into the Binary Vector pBI121 The following primers were used for amplifying the genes esat6 and cfp10 using the M. tuberculosis genomic DNA as a template: esat-upper 5′-GCTCTAGAATGACAGAGCAGCAGTGGAATTTCGCGG-3′ and esat-low 5′-CGGGATCCCTATGCGAACATCCC-3′ for the esat6 gene and for the cfp10 gene, cfp-upper 5′-GCTCTAGAATGGCAGAGATGAAGACC-3′, and cfp-low 5′-CGGGATCCGAATTCTCAGAAGCC-3′. .. After amplification, the fragments of esat6 (288 bp) and cfp10 (303 bp) genes were hydrolyzed with the restriction endonucleases Xba I and BamH I, purified using a GenElute HP Plasmid Midiprep kit (Sigma-Aldrich, United States), and ligated with the pBI121 preliminary hydrolyzed at the same sites. .. The resulting reaction mixture was used to transform E. coli strain DH10B. shows the map of the plasmid carrying the target genes (esat6 or cfp10 ).

    Article Title: Human serum albumin-mediated apoptin delivery suppresses breast cancer cell growth in vitro and in vivo
    Article Snippet: The PCR fragments were cloned into pEGFP-N1 using the T4 DNA ligase (Sigma-Aldrich; Merck Millipore). .. Subsequently, the insert site was released from pEGFP-N1 by digestion with Eco RI and Xba I restriction enzymes (Sigma-Aldrich; Merck Millipore), and PCR fragments were cloned into pEGFP-N1 using the T4 DNA ligase; the complex was termed pEGFP-N1-Apoptin, which was amplified in bacterial cells (E. coli DH5α; Takara Bio, Inc., Otsu, Japan). .. The cells were transfected with pEGFP-N1-Apoptin as follows: Following thawing of competent cells, 2 µl pEGFP-N1-Apoptin was added to 100 µl competent cells and incubated on ice for 30 min.

    Purification:

    Article Title: Oral Immunogenicity of Plant-Made Mycobacterium tuberculosis ESAT6 and CFP10
    Article Snippet: Amplification of esat6 and cfp10 Gene Sequences and Their Cloning into the Binary Vector pBI121 The following primers were used for amplifying the genes esat6 and cfp10 using the M. tuberculosis genomic DNA as a template: esat-upper 5′-GCTCTAGAATGACAGAGCAGCAGTGGAATTTCGCGG-3′ and esat-low 5′-CGGGATCCCTATGCGAACATCCC-3′ for the esat6 gene and for the cfp10 gene, cfp-upper 5′-GCTCTAGAATGGCAGAGATGAAGACC-3′, and cfp-low 5′-CGGGATCCGAATTCTCAGAAGCC-3′. .. After amplification, the fragments of esat6 (288 bp) and cfp10 (303 bp) genes were hydrolyzed with the restriction endonucleases Xba I and BamH I, purified using a GenElute HP Plasmid Midiprep kit (Sigma-Aldrich, United States), and ligated with the pBI121 preliminary hydrolyzed at the same sites. .. The resulting reaction mixture was used to transform E. coli strain DH10B. shows the map of the plasmid carrying the target genes (esat6 or cfp10 ).

    Plasmid Preparation:

    Article Title: Oral Immunogenicity of Plant-Made Mycobacterium tuberculosis ESAT6 and CFP10
    Article Snippet: Amplification of esat6 and cfp10 Gene Sequences and Their Cloning into the Binary Vector pBI121 The following primers were used for amplifying the genes esat6 and cfp10 using the M. tuberculosis genomic DNA as a template: esat-upper 5′-GCTCTAGAATGACAGAGCAGCAGTGGAATTTCGCGG-3′ and esat-low 5′-CGGGATCCCTATGCGAACATCCC-3′ for the esat6 gene and for the cfp10 gene, cfp-upper 5′-GCTCTAGAATGGCAGAGATGAAGACC-3′, and cfp-low 5′-CGGGATCCGAATTCTCAGAAGCC-3′. .. After amplification, the fragments of esat6 (288 bp) and cfp10 (303 bp) genes were hydrolyzed with the restriction endonucleases Xba I and BamH I, purified using a GenElute HP Plasmid Midiprep kit (Sigma-Aldrich, United States), and ligated with the pBI121 preliminary hydrolyzed at the same sites. .. The resulting reaction mixture was used to transform E. coli strain DH10B. shows the map of the plasmid carrying the target genes (esat6 or cfp10 ).

    Polymerase Chain Reaction:

    Article Title: Human serum albumin-mediated apoptin delivery suppresses breast cancer cell growth in vitro and in vivo
    Article Snippet: The PCR fragments were cloned into pEGFP-N1 using the T4 DNA ligase (Sigma-Aldrich; Merck Millipore). .. Subsequently, the insert site was released from pEGFP-N1 by digestion with Eco RI and Xba I restriction enzymes (Sigma-Aldrich; Merck Millipore), and PCR fragments were cloned into pEGFP-N1 using the T4 DNA ligase; the complex was termed pEGFP-N1-Apoptin, which was amplified in bacterial cells (E. coli DH5α; Takara Bio, Inc., Otsu, Japan). .. The cells were transfected with pEGFP-N1-Apoptin as follows: Following thawing of competent cells, 2 µl pEGFP-N1-Apoptin was added to 100 µl competent cells and incubated on ice for 30 min.

    Article Title: Resistance to Sri Lankan Cassava Mosaic Virus (SLCMV) in Genetically Engineered Cassava cv. KU50 through RNA Silencing
    Article Snippet: PCR amplification for the SLCMV was performed using primers specific to the SLCMV (SLCMV-5P and SLCMV-3P), whereas for npt II gene, the primers used were NPTII-5P and NPTII-3P ( ). .. For Southern blot, 15μg of the DNA was digested overnight with either Hin dIII, which cuts the T-DNA at two positions or Xba I, which cuts the T-DNA once, separated on a 0.8% agarose gel, transferred to a nylon membrane (Immobilon-Ny+ Transfer Membrane; Millipore Co, Billerica, MA, USA) and probed with SLCMV PCR-DIG labelled probe. .. The probe DNA fragment was generated from plasmid DNA pEKH2IN2SLCMV and labelled by PCR using DIG-2’-deoxyuridine 5’-triphosphate (DIG-dUTP) in the sense and antisense direction according to the supplier’s instruction (PCR DIG probe synthesis kit, Roche Diagnostic GmbH, Boehringer Mannheim, Germany).

    Southern Blot:

    Article Title: Resistance to Sri Lankan Cassava Mosaic Virus (SLCMV) in Genetically Engineered Cassava cv. KU50 through RNA Silencing
    Article Snippet: PCR amplification for the SLCMV was performed using primers specific to the SLCMV (SLCMV-5P and SLCMV-3P), whereas for npt II gene, the primers used were NPTII-5P and NPTII-3P ( ). .. For Southern blot, 15μg of the DNA was digested overnight with either Hin dIII, which cuts the T-DNA at two positions or Xba I, which cuts the T-DNA once, separated on a 0.8% agarose gel, transferred to a nylon membrane (Immobilon-Ny+ Transfer Membrane; Millipore Co, Billerica, MA, USA) and probed with SLCMV PCR-DIG labelled probe. .. The probe DNA fragment was generated from plasmid DNA pEKH2IN2SLCMV and labelled by PCR using DIG-2’-deoxyuridine 5’-triphosphate (DIG-dUTP) in the sense and antisense direction according to the supplier’s instruction (PCR DIG probe synthesis kit, Roche Diagnostic GmbH, Boehringer Mannheim, Germany).

    Agarose Gel Electrophoresis:

    Article Title: Resistance to Sri Lankan Cassava Mosaic Virus (SLCMV) in Genetically Engineered Cassava cv. KU50 through RNA Silencing
    Article Snippet: PCR amplification for the SLCMV was performed using primers specific to the SLCMV (SLCMV-5P and SLCMV-3P), whereas for npt II gene, the primers used were NPTII-5P and NPTII-3P ( ). .. For Southern blot, 15μg of the DNA was digested overnight with either Hin dIII, which cuts the T-DNA at two positions or Xba I, which cuts the T-DNA once, separated on a 0.8% agarose gel, transferred to a nylon membrane (Immobilon-Ny+ Transfer Membrane; Millipore Co, Billerica, MA, USA) and probed with SLCMV PCR-DIG labelled probe. .. The probe DNA fragment was generated from plasmid DNA pEKH2IN2SLCMV and labelled by PCR using DIG-2’-deoxyuridine 5’-triphosphate (DIG-dUTP) in the sense and antisense direction according to the supplier’s instruction (PCR DIG probe synthesis kit, Roche Diagnostic GmbH, Boehringer Mannheim, Germany).

    other:

    Article Title: Whole-Genome Shotgun Optical Mapping of Rhodospirillum rubrum
    Article Snippet: Surface properties were assayed by digesting lambda DASH II bacteriophage DNA with 40 units of XbaI, HindIII, and NheI diluted in 200 μl of digestion buffer with 0.2% Triton X-100 (Sigma, St. Louis, MO) at 37°C to determine optimal digestion times, which ranged from 30 to 120 min.

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    • xba i  (Millipore)
    93
    Millipore xba i
    Dendrogram based on <t>Xba</t> I PFGE types identified among 31 E. coli strains. The data were sorted by the UPGMA method. AMP-ampicillin, TET-tetracycline, SUT- trimethoprim/sulfamethoxazole, CLO-chloramphenicol, CFL-cephalothin, AMC-amoxicillin/clavulanic acid, LVX- levofloxacin, CIP-ciprofloxacin, GEN-gentamicin, CRO-ceftriaxone, CTX-cefotaxime. *Isolates that show intermediate susceptibility to cephalotin.
    Xba I, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/xba i/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    xba i - by Bioz Stars, 2021-03
    93/100 stars
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    Dendrogram based on Xba I PFGE types identified among 31 E. coli strains. The data were sorted by the UPGMA method. AMP-ampicillin, TET-tetracycline, SUT- trimethoprim/sulfamethoxazole, CLO-chloramphenicol, CFL-cephalothin, AMC-amoxicillin/clavulanic acid, LVX- levofloxacin, CIP-ciprofloxacin, GEN-gentamicin, CRO-ceftriaxone, CTX-cefotaxime. *Isolates that show intermediate susceptibility to cephalotin.

    Journal: Brazilian Journal of Microbiology

    Article Title: Antimicrobial resistance and genetic diversity of Escherichiacoli isolated from humans and foods

    doi: 10.1590/S1517-838246420130874

    Figure Lengend Snippet: Dendrogram based on Xba I PFGE types identified among 31 E. coli strains. The data were sorted by the UPGMA method. AMP-ampicillin, TET-tetracycline, SUT- trimethoprim/sulfamethoxazole, CLO-chloramphenicol, CFL-cephalothin, AMC-amoxicillin/clavulanic acid, LVX- levofloxacin, CIP-ciprofloxacin, GEN-gentamicin, CRO-ceftriaxone, CTX-cefotaxime. *Isolates that show intermediate susceptibility to cephalotin.

    Article Snippet: Data Analysis The Xba I (Sigma-Aldrich, USA) fingerprints were analyzed using GelCompar II software (Applied Maths, Kortrijk, Belgium).

    Techniques:

    Luciferase activity of J6CF backbone replicons containing substitutions in the NS5B region. (A) Schematic structures of wt JFH-1 and J6CF constructs and the chimeric subgenomic replicons containing a J6CF backbone. The restriction enzyme recognition sites used for the construction of plasmids are indicated. C , Cla I; E , Eco T22I; B , Bsr GI; S , Stu I; X , Xba I; wt, wild-type. (B) Schematic diagram of the mutations introduced into J6/N3H+3′UTR-JFH1-Luc and J6/N3H+3′UTR-JFH1. (C) Replication activity of J6CF-based replicons. Subgenomic RNA was synthesized in vitro from wild-type or chimeric replicon constructs. Transcribed subgenomic RNA (5 µg) was then electroporated into HuH-7 cells and the cells harvested at 4, 24, and 48 h after transfection. The harvested cells were lysed, and the luciferase activity in the cell lysates was measured. The assays were performed three times independently and the results expressed as luciferase activities (RLU). Each value was corrected for transfection efficiency as determined by measuring the luciferase activity 4 h after transfection. Data are presented as the mean ± standard deviation for luciferase activity at 24 h (white bars) and 48 h (gray bars) after transfection. Asterisks indicate significant differences relative to the replication activity of J6/N3H+N5BX-JFH1 (p

    Journal: PLoS Pathogens

    Article Title: RNA Polymerase Activity and Specific RNA Structure Are Required for Efficient HCV Replication in Cultured Cells

    doi: 10.1371/journal.ppat.1000885

    Figure Lengend Snippet: Luciferase activity of J6CF backbone replicons containing substitutions in the NS5B region. (A) Schematic structures of wt JFH-1 and J6CF constructs and the chimeric subgenomic replicons containing a J6CF backbone. The restriction enzyme recognition sites used for the construction of plasmids are indicated. C , Cla I; E , Eco T22I; B , Bsr GI; S , Stu I; X , Xba I; wt, wild-type. (B) Schematic diagram of the mutations introduced into J6/N3H+3′UTR-JFH1-Luc and J6/N3H+3′UTR-JFH1. (C) Replication activity of J6CF-based replicons. Subgenomic RNA was synthesized in vitro from wild-type or chimeric replicon constructs. Transcribed subgenomic RNA (5 µg) was then electroporated into HuH-7 cells and the cells harvested at 4, 24, and 48 h after transfection. The harvested cells were lysed, and the luciferase activity in the cell lysates was measured. The assays were performed three times independently and the results expressed as luciferase activities (RLU). Each value was corrected for transfection efficiency as determined by measuring the luciferase activity 4 h after transfection. Data are presented as the mean ± standard deviation for luciferase activity at 24 h (white bars) and 48 h (gray bars) after transfection. Asterisks indicate significant differences relative to the replication activity of J6/N3H+N5BX-JFH1 (p

    Article Snippet: Following digestion with Xba I and Xho I, DNA fragments were cloned into the Nhe I and Xho I sites of pET21b (Novagen, Madison, WI), resulting in pET21bHCVJFH-1RdRpwt and pET21bHCVJ6CFRdRpwt. pET21bHCVJFH-1RdRpwt and pET21bHCVJ6CFRdRpwt were then digested with Xba I and Xho I and the RdRP fragments cloned into the same restriction sites of pET28a, resulting in pET21(KM)JFH-1RdRpwt and pET21(KM)J6CFRdRpwt, respectively.

    Techniques: Luciferase, Activity Assay, Construct, Synthesized, In Vitro, Transfection, Standard Deviation

    Analysis of transient replication of genomic chimeric HCV RNA. (A) The structure of full-length chimeric HCV RNA. Each chimeric full-length construct was prepared by the replacement of the indicated restricted fragments. The restriction enzyme recognition sites used for the plasmid constructions are indicated. C , Cla I; E , Eco T22I; B , Bsr GI; S , Stu I; X , Xba I; wt, wild-type. (B) HCV core protein production in the culture medium from RNA-transfected cells. Transcribed wild-type or chimeric full-length HCV RNA (10 µg) was transfected into Huh-7.5.1 cells. Culture medium was harvested at 4, 24, 48, and 72 h after transfection. The amount of core protein in the harvested culture medium was measured using a HCV core chemiluminescence enzyme immunoassay (Lumipulse II HCV core assay). The assays were performed three times independently, and the data are presented as the mean ± standard deviation. Values in the right panel represent the relative core values against J6/N3H+N5BX-JFH1 at 72 h after transfection and infectious titers of the media from chimeric HCV RNA-transfected cells at 72h after transfection determined using Huh7.5.1 cells. SNF, A450S+S455N+Y561F; SKF, A450S+R517K+Y561F; SNKF, A450S+ S455N+R517K+Y561F.

    Journal: PLoS Pathogens

    Article Title: RNA Polymerase Activity and Specific RNA Structure Are Required for Efficient HCV Replication in Cultured Cells

    doi: 10.1371/journal.ppat.1000885

    Figure Lengend Snippet: Analysis of transient replication of genomic chimeric HCV RNA. (A) The structure of full-length chimeric HCV RNA. Each chimeric full-length construct was prepared by the replacement of the indicated restricted fragments. The restriction enzyme recognition sites used for the plasmid constructions are indicated. C , Cla I; E , Eco T22I; B , Bsr GI; S , Stu I; X , Xba I; wt, wild-type. (B) HCV core protein production in the culture medium from RNA-transfected cells. Transcribed wild-type or chimeric full-length HCV RNA (10 µg) was transfected into Huh-7.5.1 cells. Culture medium was harvested at 4, 24, 48, and 72 h after transfection. The amount of core protein in the harvested culture medium was measured using a HCV core chemiluminescence enzyme immunoassay (Lumipulse II HCV core assay). The assays were performed three times independently, and the data are presented as the mean ± standard deviation. Values in the right panel represent the relative core values against J6/N3H+N5BX-JFH1 at 72 h after transfection and infectious titers of the media from chimeric HCV RNA-transfected cells at 72h after transfection determined using Huh7.5.1 cells. SNF, A450S+S455N+Y561F; SKF, A450S+R517K+Y561F; SNKF, A450S+ S455N+R517K+Y561F.

    Article Snippet: Following digestion with Xba I and Xho I, DNA fragments were cloned into the Nhe I and Xho I sites of pET21b (Novagen, Madison, WI), resulting in pET21bHCVJFH-1RdRpwt and pET21bHCVJ6CFRdRpwt. pET21bHCVJFH-1RdRpwt and pET21bHCVJ6CFRdRpwt were then digested with Xba I and Xho I and the RdRP fragments cloned into the same restriction sites of pET28a, resulting in pET21(KM)JFH-1RdRpwt and pET21(KM)J6CFRdRpwt, respectively.

    Techniques: Construct, Plasmid Preparation, Transfection, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Comparison of digestion pattern of eDNA and genomic DNA (by DNase I and restriction enzymes). Agarose gel (0.8%) showing Lane 1: Genomic DNA, Lane 2: Digestion of genomic DNA with DNase I, Lane 3: Digestion of genomic DNA with restriction enzymes ( Bam HI, Eco RI, Hin dIII, Pst I, Xba I, Xho I), Lane M: Molecular weight marker ( λ phage genome Eco RI /Hin dIII digest), Lane 4: Digestion of eDNA with restriction enzymes ( Bam HI, Eco RI, Hin dIII, Pst I, Xba I, Xho I), Lane 5: Digestion of eDNA with DNase I, Lane 6: Purified eDNA.

    Journal: The Scientific World Journal

    Article Title: Characterization of eDNA from the Clinical Strain Acinetobacter baumannii AIIMS 7 and Its Role in Biofilm Formation

    doi: 10.1100/2012/973436

    Figure Lengend Snippet: Comparison of digestion pattern of eDNA and genomic DNA (by DNase I and restriction enzymes). Agarose gel (0.8%) showing Lane 1: Genomic DNA, Lane 2: Digestion of genomic DNA with DNase I, Lane 3: Digestion of genomic DNA with restriction enzymes ( Bam HI, Eco RI, Hin dIII, Pst I, Xba I, Xho I), Lane M: Molecular weight marker ( λ phage genome Eco RI /Hin dIII digest), Lane 4: Digestion of eDNA with restriction enzymes ( Bam HI, Eco RI, Hin dIII, Pst I, Xba I, Xho I), Lane 5: Digestion of eDNA with DNase I, Lane 6: Purified eDNA.

    Article Snippet: Digestion of eDNA by DNase I and Restriction Enzymes To check the digestion pattern of eDNA and compare with genomic DNA, eDNA was digested with six different restriction enzymes Bam HI, Eco RI, Hin dIII, Pst I, Xba I, and Xho I (Sigma Aldrich).

    Techniques: Agarose Gel Electrophoresis, Molecular Weight, Marker, Purification

    Successful establishment of the human serum albumin-p olyethylenimine-pcDNA-Apoptin expression vector was confirmed by 1% agarose gel electrophoresis. Lane 1, DL15000 DNA marker. Lanes 2 and 3, pcDNA-Apoptin was digested by Eco RI and Xba I restriction enzymes, and produced two fragments: apoptin at 376 bp and p-enhanced green fluorescent protein (EGFP)-N1 at 5,000 bp. Lanes 4 and 5, pEGFP-N1 was digested by Eco RI and Xba I restriction enzymes, and produced only one fragment at 5,000 bp. Lanes 6 and 7, the pEGFP-N1 plasmid.

    Journal: Oncology Letters

    Article Title: Human serum albumin-mediated apoptin delivery suppresses breast cancer cell growth in vitro and in vivo

    doi: 10.3892/ol.2016.5470

    Figure Lengend Snippet: Successful establishment of the human serum albumin-p olyethylenimine-pcDNA-Apoptin expression vector was confirmed by 1% agarose gel electrophoresis. Lane 1, DL15000 DNA marker. Lanes 2 and 3, pcDNA-Apoptin was digested by Eco RI and Xba I restriction enzymes, and produced two fragments: apoptin at 376 bp and p-enhanced green fluorescent protein (EGFP)-N1 at 5,000 bp. Lanes 4 and 5, pEGFP-N1 was digested by Eco RI and Xba I restriction enzymes, and produced only one fragment at 5,000 bp. Lanes 6 and 7, the pEGFP-N1 plasmid.

    Article Snippet: Subsequently, the insert site was released from pEGFP-N1 by digestion with Eco RI and Xba I restriction enzymes (Sigma-Aldrich; Merck Millipore), and PCR fragments were cloned into pEGFP-N1 using the T4 DNA ligase; the complex was termed pEGFP-N1-Apoptin, which was amplified in bacterial cells (E. coli DH5α; Takara Bio, Inc., Otsu, Japan).

    Techniques: Expressing, Plasmid Preparation, Agarose Gel Electrophoresis, Marker, Produced