kpn i  (Thermo Fisher)


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    Name:
    KpnI
    Description:
    5 G G T A C ↓C 3 3 C ↑C A T G G 5 Thermo Scientific KpnI restriction enzyme recognizes GGTAC C sites and cuts best at 37°C in its own unique buffer See Reaction Conditions for Restriction Enzymes for a table of enzyme activity conditions for double digestion and heat inactivation for this and other restriction enzymes Isoschizomers Asp718I Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes optimized to work in one of the buffers of the Five Buffer System In addition the universal Tango buffer is provided for convenience in double digestions All of the enzymes exhibit 100 activity in the recommended buffer and reaction conditions To ensure consistent performance Thermo Scientific restriction enzyme reaction buffers contain premixed BSA which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations Features• Superior quality stringent quality control and industry leading manufacturing process• Convenient color coded Five Buffer System• Includes universal Tango buffer for double digestions• BSA premixed in reaction buffers• Wide selection of restriction endonuclease specificitiesApplications• Molecular cloning• Restriction site mapping• Genotyping• Southern blotting• Restriction fragment length polymorphism RFLP • SNPNote For methylation sensitivity refer to product specifications
    Catalog Number:
    er0523
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    None
    Applications:
    Cloning|Restriction Enzyme Cloning
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher kpn i
    End configuration of the <t>DNA</t> substrates used in this study. The diagram presents the DNA ends created by restriction of M13 mp18 derivatives with Xma I and Mlu I (XM), Sma I (S) or Aat II and <t>Kpn</t> I (AK). Sites of homology to the protruding DNA ends are shown in bold.
    5 G G T A C ↓C 3 3 C ↑C A T G G 5 Thermo Scientific KpnI restriction enzyme recognizes GGTAC C sites and cuts best at 37°C in its own unique buffer See Reaction Conditions for Restriction Enzymes for a table of enzyme activity conditions for double digestion and heat inactivation for this and other restriction enzymes Isoschizomers Asp718I Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes optimized to work in one of the buffers of the Five Buffer System In addition the universal Tango buffer is provided for convenience in double digestions All of the enzymes exhibit 100 activity in the recommended buffer and reaction conditions To ensure consistent performance Thermo Scientific restriction enzyme reaction buffers contain premixed BSA which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations Features• Superior quality stringent quality control and industry leading manufacturing process• Convenient color coded Five Buffer System• Includes universal Tango buffer for double digestions• BSA premixed in reaction buffers• Wide selection of restriction endonuclease specificitiesApplications• Molecular cloning• Restriction site mapping• Genotyping• Southern blotting• Restriction fragment length polymorphism RFLP • SNPNote For methylation sensitivity refer to product specifications
    https://www.bioz.com/result/kpn i/product/Thermo Fisher
    Average 99 stars, based on 329 article reviews
    Price from $9.99 to $1999.99
    kpn i - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "The role of DNA polymerase activity in human non-homologous end joining"

    Article Title: The role of DNA polymerase activity in human non-homologous end joining

    Journal: Nucleic Acids Research

    doi:

    End configuration of the DNA substrates used in this study. The diagram presents the DNA ends created by restriction of M13 mp18 derivatives with Xma I and Mlu I (XM), Sma I (S) or Aat II and Kpn I (AK). Sites of homology to the protruding DNA ends are shown in bold.
    Figure Legend Snippet: End configuration of the DNA substrates used in this study. The diagram presents the DNA ends created by restriction of M13 mp18 derivatives with Xma I and Mlu I (XM), Sma I (S) or Aat II and Kpn I (AK). Sites of homology to the protruding DNA ends are shown in bold.

    Techniques Used:

    2) Product Images from "Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein"

    Article Title: Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein

    Journal: Retrovirology

    doi: 10.1186/1742-4690-9-17

    Schematic construction of the ankyrin repeat library using the pHDiEx acceptor vector . (A) , The mono-ankyrin microgenes were polymerized by insertion/ligation to pH. DiEx double digested by Bsm BI and Kpn I. The construction was subsequently digested by Bsp MI and recircularised by intramolecular ligation. This resulted in the substitution of the region containing the Rep cloning sites by the ankyrin repeats. The number of repeats differed from one clone to another, and usually ranged from 2 to 6 repeats. (B) , Detailed sequences of the different DNA regions. Abbreviations: T7p, T7 promoter; pLac/RBS, lactose promoter and Ribosome binding site; DsbA ss, DsbA signal sequence; St2, Strep-Tag2 tag; H6, hexa-histidine tag.
    Figure Legend Snippet: Schematic construction of the ankyrin repeat library using the pHDiEx acceptor vector . (A) , The mono-ankyrin microgenes were polymerized by insertion/ligation to pH. DiEx double digested by Bsm BI and Kpn I. The construction was subsequently digested by Bsp MI and recircularised by intramolecular ligation. This resulted in the substitution of the region containing the Rep cloning sites by the ankyrin repeats. The number of repeats differed from one clone to another, and usually ranged from 2 to 6 repeats. (B) , Detailed sequences of the different DNA regions. Abbreviations: T7p, T7 promoter; pLac/RBS, lactose promoter and Ribosome binding site; DsbA ss, DsbA signal sequence; St2, Strep-Tag2 tag; H6, hexa-histidine tag.

    Techniques Used: Plasmid Preparation, Ligation, Clone Assay, Binding Assay, Sequencing

    3) Product Images from "Construction and Characterization of a 1,3-Propanediol Operon"

    Article Title: Construction and Characterization of a 1,3-Propanediol Operon

    Journal: Applied and Environmental Microbiology

    doi:

    Construction of pTC48 as outlined in Materials and Methods. Restriction enzyme recognition sites are abbreviated as follows: H, Hin dIII; K, Kpn I; Nd, Nde I; Nh, Nhe I; No, Not I; P, Pst I; Sc, Sac I; and Sl, Sal I. Brackets indicate the combination of two fragments of DNA by cleavage at the restriction sites shown in the bracket and subsequent ligation to form the product shown beneath the bracket.
    Figure Legend Snippet: Construction of pTC48 as outlined in Materials and Methods. Restriction enzyme recognition sites are abbreviated as follows: H, Hin dIII; K, Kpn I; Nd, Nde I; Nh, Nhe I; No, Not I; P, Pst I; Sc, Sac I; and Sl, Sal I. Brackets indicate the combination of two fragments of DNA by cleavage at the restriction sites shown in the bracket and subsequent ligation to form the product shown beneath the bracket.

    Techniques Used: Ligation

    4) Product Images from "Mucosal and systemic immune responses elicited by recombinant Lactococcus lactis expressing a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis"

    Article Title: Mucosal and systemic immune responses elicited by recombinant Lactococcus lactis expressing a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis

    Journal: Microbial Pathogenesis

    doi: 10.1016/j.micpath.2018.05.008

    Identification of the recombinant plasmid pNZ8149-SECF1S1 by PCR and restriction enzyme analysis (a) PCR amplification of F1S1gene. Lane A: DNA ladder; Lanes B and C: PCR products of the F1S1 gene amplification D: No template control. (b) Restriction analysis of the recombinant plasmid pNZ8149-SECF1S1. Lane A: DNA ladder; Lane B: pNZ8149-SECF1S1 double-digested with Kpn I and Xba I; C: pNZ8149-SECF1S1 digested with Kpn I.
    Figure Legend Snippet: Identification of the recombinant plasmid pNZ8149-SECF1S1 by PCR and restriction enzyme analysis (a) PCR amplification of F1S1gene. Lane A: DNA ladder; Lanes B and C: PCR products of the F1S1 gene amplification D: No template control. (b) Restriction analysis of the recombinant plasmid pNZ8149-SECF1S1. Lane A: DNA ladder; Lane B: pNZ8149-SECF1S1 double-digested with Kpn I and Xba I; C: pNZ8149-SECF1S1 digested with Kpn I.

    Techniques Used: Recombinant, Plasmid Preparation, Polymerase Chain Reaction, Amplification

    5) Product Images from "Rapid and Efficient Purification of Drosophila Homeodomain Transcription Factors for Biophysical Characterization"

    Article Title: Rapid and Efficient Purification of Drosophila Homeodomain Transcription Factors for Biophysical Characterization

    Journal: Protein expression and purification

    doi: 10.1016/j.pep.2019.02.001

    Cloning and sequence alignment of D. melanogaster HDs for bacterial overexpression. Vector map (A) and sequence map (B) of HisSUMO-HD constructs. Each of the HDs for FTZ, ABD-A, ABD-B, ANTP, and UBX were cloned into a T7-inducible HisSUMO vector between Kpn I and Xba I restriction sites. (C) Sequence alignment of overexpressed D. melanogaster ]. Only the five HDs studied in this paper are shown. Highly conserved residues within similar physicochemical properties in all 107 HDs analysed are shown (red – positive, blue – negative, yellow – polar, green – hydrophobic). Absolutely conserved residues and residues with greater than 70% similarity are labelled with a star or cross, respectively.
    Figure Legend Snippet: Cloning and sequence alignment of D. melanogaster HDs for bacterial overexpression. Vector map (A) and sequence map (B) of HisSUMO-HD constructs. Each of the HDs for FTZ, ABD-A, ABD-B, ANTP, and UBX were cloned into a T7-inducible HisSUMO vector between Kpn I and Xba I restriction sites. (C) Sequence alignment of overexpressed D. melanogaster ]. Only the five HDs studied in this paper are shown. Highly conserved residues within similar physicochemical properties in all 107 HDs analysed are shown (red – positive, blue – negative, yellow – polar, green – hydrophobic). Absolutely conserved residues and residues with greater than 70% similarity are labelled with a star or cross, respectively.

    Techniques Used: Clone Assay, Sequencing, Over Expression, Plasmid Preparation, Construct

    6) Product Images from "Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein"

    Article Title: Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein

    Journal: Retrovirology

    doi: 10.1186/1742-4690-9-17

    Schematic construction of the ankyrin repeat library using the pHDiEx acceptor vector . (A) , The mono-ankyrin microgenes were polymerized by insertion/ligation to pH. DiEx double digested by Bsm BI and Kpn I. The construction was subsequently digested by Bsp MI and recircularised by intramolecular ligation. This resulted in the substitution of the region containing the Rep cloning sites by the ankyrin repeats. The number of repeats differed from one clone to another, and usually ranged from 2 to 6 repeats. (B) , Detailed sequences of the different DNA regions. Abbreviations: T7p, T7 promoter; pLac/RBS, lactose promoter and Ribosome binding site; DsbA ss, DsbA signal sequence; St2, Strep-Tag2 tag; H6, hexa-histidine tag.
    Figure Legend Snippet: Schematic construction of the ankyrin repeat library using the pHDiEx acceptor vector . (A) , The mono-ankyrin microgenes were polymerized by insertion/ligation to pH. DiEx double digested by Bsm BI and Kpn I. The construction was subsequently digested by Bsp MI and recircularised by intramolecular ligation. This resulted in the substitution of the region containing the Rep cloning sites by the ankyrin repeats. The number of repeats differed from one clone to another, and usually ranged from 2 to 6 repeats. (B) , Detailed sequences of the different DNA regions. Abbreviations: T7p, T7 promoter; pLac/RBS, lactose promoter and Ribosome binding site; DsbA ss, DsbA signal sequence; St2, Strep-Tag2 tag; H6, hexa-histidine tag.

    Techniques Used: Plasmid Preparation, Ligation, Clone Assay, Binding Assay, Sequencing

    7) Product Images from "Unusual organization of a developmentally regulated mitochondrial RNA polymerase (TBMTRNAP) gene in Trypanosoma brucei"

    Article Title: Unusual organization of a developmentally regulated mitochondrial RNA polymerase (TBMTRNAP) gene in Trypanosoma brucei

    Journal: Gene

    doi:

    (A) Restriction map of the TBMTRNAP locus and cloning strategy. Numbers are given relative to the first nt in the TBMTRNAP ORF. The TBMTRNAP ORF is represented by a long rectangle within which the degenerate PCR product is boxed. The bold lines above the restriction map indicate the subcloned P1 fragments. Arrows beneath the map indicate the cDNA clones. The 5′ and 3′ UTRs are represented by lines extending from the TBMTRNAP ORF. SAS indicates the splice acceptor site. The multiple polyadenylation sites are indicated by the downward arrows. Relative positions of the primers used in the 5′ RTPCR experiments are indicated by arrowheads above the map. (B) Southern blot analysis of the T. brucei genomic DNA. DNA (10 µg) was digested with Bam HI (lane 1), Eco RI (lane 2), Kpn I (lane 4), Bam HI + Eco RI (lane 3), Kpn I + Bam HI (lane 5), and Kpn I + Eco RI (lane 6), separated on a 0.75% gel, transferred to Nytran, and hybridized with a riboprobe spanning the Kpn I site at 1907 and the Bam HI site at 3759.
    Figure Legend Snippet: (A) Restriction map of the TBMTRNAP locus and cloning strategy. Numbers are given relative to the first nt in the TBMTRNAP ORF. The TBMTRNAP ORF is represented by a long rectangle within which the degenerate PCR product is boxed. The bold lines above the restriction map indicate the subcloned P1 fragments. Arrows beneath the map indicate the cDNA clones. The 5′ and 3′ UTRs are represented by lines extending from the TBMTRNAP ORF. SAS indicates the splice acceptor site. The multiple polyadenylation sites are indicated by the downward arrows. Relative positions of the primers used in the 5′ RTPCR experiments are indicated by arrowheads above the map. (B) Southern blot analysis of the T. brucei genomic DNA. DNA (10 µg) was digested with Bam HI (lane 1), Eco RI (lane 2), Kpn I (lane 4), Bam HI + Eco RI (lane 3), Kpn I + Bam HI (lane 5), and Kpn I + Eco RI (lane 6), separated on a 0.75% gel, transferred to Nytran, and hybridized with a riboprobe spanning the Kpn I site at 1907 and the Bam HI site at 3759.

    Techniques Used: Clone Assay, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Southern Blot

    8) Product Images from "Immune Protection of Rhoptry Protein 21 (ROP21) of Toxoplasma gondii as a DNA Vaccine Against Toxoplasmosis"

    Article Title: Immune Protection of Rhoptry Protein 21 (ROP21) of Toxoplasma gondii as a DNA Vaccine Against Toxoplasmosis

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.00909

    Agarose gel electrophoresis of TgROP21 ORF and identification of recombinant plasmid pVAX-TgROP21 digested by Kpn I and ECO RI. (A) (Lane M) DNA molecular weight marker DL 2000 (ordinate values in bp); (Lane 1) the ORF of TgROP21. (B) (Lane M) DNA molecular weight marker DL 2000 (ordinate values in bp); (Lane 1) the recombinant plasmid pVAX-TgROP21 digested by Kpn I and ECO RI; (Lane 2) the recombinant plasmid pVAX-TgROP21; (Lane 3) the plasmid of pVAXI vector digested by Kpn I and ECO RI.
    Figure Legend Snippet: Agarose gel electrophoresis of TgROP21 ORF and identification of recombinant plasmid pVAX-TgROP21 digested by Kpn I and ECO RI. (A) (Lane M) DNA molecular weight marker DL 2000 (ordinate values in bp); (Lane 1) the ORF of TgROP21. (B) (Lane M) DNA molecular weight marker DL 2000 (ordinate values in bp); (Lane 1) the recombinant plasmid pVAX-TgROP21 digested by Kpn I and ECO RI; (Lane 2) the recombinant plasmid pVAX-TgROP21; (Lane 3) the plasmid of pVAXI vector digested by Kpn I and ECO RI.

    Techniques Used: Agarose Gel Electrophoresis, Recombinant, Plasmid Preparation, Molecular Weight, Marker

    9) Product Images from "Identification of Critical Elements for Regulation of Inorganic Pyrophosphatase (PPA1) in MCF7 Breast Cancer Cells"

    Article Title: Identification of Critical Elements for Regulation of Inorganic Pyrophosphatase (PPA1) in MCF7 Breast Cancer Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0124864

    Identification of PPA1 minimal promoter. (A) Determination of transcription start site of PPA1 gene. Total RNA was isolated from MCF7 cells and analyzed by primer extension analysis using a primer complementary to the -20 to -1 region of exon 1 of PPA1 gene. The sequencing reaction was set using same primer as in primer extension. Both the sequencing reaction products (A, T, G and C) and primer extension product (PE) were run on a 7.5% urea-polyacrylamide gel. (B) Schematic representation of 5’-deletion constructs of PPA1 promoter. All the deletion constructs were generated by PCR using pGL3-1217 construct as template and cloned into the Kpn-I and Xho-I restriction site of pGL3-Basic vector. (C) Promoter activity of all the 5’-deleted promoter constructs was determined by Dual-Luciferase assay. MCF7 cells were transfected with promoter constructs and after 24 h luciferase assay was carried out using Dual-Luciferase assay kit (Promega). Each transfection was conducted in triplicate and the results are represented as mean ± S.E.M.
    Figure Legend Snippet: Identification of PPA1 minimal promoter. (A) Determination of transcription start site of PPA1 gene. Total RNA was isolated from MCF7 cells and analyzed by primer extension analysis using a primer complementary to the -20 to -1 region of exon 1 of PPA1 gene. The sequencing reaction was set using same primer as in primer extension. Both the sequencing reaction products (A, T, G and C) and primer extension product (PE) were run on a 7.5% urea-polyacrylamide gel. (B) Schematic representation of 5’-deletion constructs of PPA1 promoter. All the deletion constructs were generated by PCR using pGL3-1217 construct as template and cloned into the Kpn-I and Xho-I restriction site of pGL3-Basic vector. (C) Promoter activity of all the 5’-deleted promoter constructs was determined by Dual-Luciferase assay. MCF7 cells were transfected with promoter constructs and after 24 h luciferase assay was carried out using Dual-Luciferase assay kit (Promega). Each transfection was conducted in triplicate and the results are represented as mean ± S.E.M.

    Techniques Used: Isolation, Sequencing, Construct, Generated, Polymerase Chain Reaction, Clone Assay, Plasmid Preparation, Activity Assay, Luciferase, Transfection

    10) Product Images from "Immune Protection of Rhoptry Protein 21 (ROP21) of Toxoplasma gondii as a DNA Vaccine Against Toxoplasmosis"

    Article Title: Immune Protection of Rhoptry Protein 21 (ROP21) of Toxoplasma gondii as a DNA Vaccine Against Toxoplasmosis

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.00909

    Agarose gel electrophoresis of TgROP21 ORF and identification of recombinant plasmid pVAX-TgROP21 digested by Kpn I and ECO RI. (A) (Lane M) DNA molecular weight marker DL 2000 (ordinate values in bp); (Lane 1) the ORF of TgROP21. (B) (Lane M) DNA molecular weight marker DL 2000 (ordinate values in bp); (Lane 1) the recombinant plasmid pVAX-TgROP21 digested by Kpn I and ECO RI; (Lane 2) the recombinant plasmid pVAX-TgROP21; (Lane 3) the plasmid of pVAXI vector digested by Kpn I and ECO RI.
    Figure Legend Snippet: Agarose gel electrophoresis of TgROP21 ORF and identification of recombinant plasmid pVAX-TgROP21 digested by Kpn I and ECO RI. (A) (Lane M) DNA molecular weight marker DL 2000 (ordinate values in bp); (Lane 1) the ORF of TgROP21. (B) (Lane M) DNA molecular weight marker DL 2000 (ordinate values in bp); (Lane 1) the recombinant plasmid pVAX-TgROP21 digested by Kpn I and ECO RI; (Lane 2) the recombinant plasmid pVAX-TgROP21; (Lane 3) the plasmid of pVAXI vector digested by Kpn I and ECO RI.

    Techniques Used: Agarose Gel Electrophoresis, Recombinant, Plasmid Preparation, Molecular Weight, Marker

    Related Articles

    Clone Assay:

    Article Title: Construction and Characterization of a 1,3-Propanediol Operon
    Article Snippet: .. This product was cut with Kpn I and Pst I and inserted into pSE380 (Invitrogen) cut with the same two enzymes to give pTC35. pTC35 was cut with Pst I and Sac I, both originally in the multiple cloning site of pSE380, so that the remainder of the dhaT gene (the Pst I- Sac I fragment of pTC3) could be inserted. .. The resulting plasmid contained the full dhaT gene under the control of the trc promoter.

    Article Title: Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
    Article Snippet: .. The PCR products of the second round were treated with Kpn I and Xho I (Fermentas) and cloned into corresponding sites of the pCEP4 vector. .. The sequence of AnkGAG 1D4-GFP and AnkA3 2D3-GFP, as well as all our other constructs, was verified by standard DNA sequencing.

    Synthesized:

    Article Title: Mucosal and systemic immune responses elicited by recombinant Lactococcus lactis expressing a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis
    Article Snippet: .. The synthesized fragment (SECF1S1) was digested with Kpn I and Xba I restriction endonucleases (Thermo Fisher Scientific) and was inserted into the corresponding sites of the expression vector pNZ8149, giving rise to pNZ-SECF1S1. ..

    Sequencing:

    Article Title: PepGMV Rep-Protein Expression in Mammalian Cells
    Article Snippet: .. The insert presence was confirmed by double digestion with SpeI and KpnI enzymes (Fermentas).Gene correct orientation was confirmed by sequencing (Genetic Analyzer 3130, Applied Biosystems). .. Cell Culture and Transfection 3T3L1 mice fibroblast cells (ATCC No. CL-173) were grown in Dubelco’s Modified Eagle Medium (DMEM, Gibco) supplemented with penicillin (62.1 mg/L; Sigma), streptomycin (100 mg/L; Sigma), anphotericin (250 µg/L; Gibco), and 5% of calf serum (Biowest).

    Expressing:

    Article Title: Mucosal and systemic immune responses elicited by recombinant Lactococcus lactis expressing a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis
    Article Snippet: .. The synthesized fragment (SECF1S1) was digested with Kpn I and Xba I restriction endonucleases (Thermo Fisher Scientific) and was inserted into the corresponding sites of the expression vector pNZ8149, giving rise to pNZ-SECF1S1. ..

    Polymerase Chain Reaction:

    Article Title: Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
    Article Snippet: .. The PCR products of the second round were treated with Kpn I and Xho I (Fermentas) and cloned into corresponding sites of the pCEP4 vector. .. The sequence of AnkGAG 1D4-GFP and AnkA3 2D3-GFP, as well as all our other constructs, was verified by standard DNA sequencing.

    Plasmid Preparation:

    Article Title: Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
    Article Snippet: .. The PCR products of the second round were treated with Kpn I and Xho I (Fermentas) and cloned into corresponding sites of the pCEP4 vector. .. The sequence of AnkGAG 1D4-GFP and AnkA3 2D3-GFP, as well as all our other constructs, was verified by standard DNA sequencing.

    Article Title: Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
    Article Snippet: .. This vector was first cleaved with Bsm B I and Kpn I (Fermentas) to generate the cohesive ends compatible with ankyrin repeats microgenes. .. The Kpn I cleavage, although not strictly necessary, was used to minimize the vector recircularization which would compete with ankyrin-repeat polymerisation.

    Article Title: PepGMV Rep-Protein Expression in Mammalian Cells
    Article Snippet: .. The pTracer™-SV40 vector (Invitrogen) was double digested with SpeI and KpnI enzymes (Fermentas) in order to linearize it and clone the Rep gene. .. Insert and vector purification was performed using the Silica Bead DNA Gel Extraction kit (Fermentas) and ligation reaction was done using the ligase T4 (Invitrogen) and a 4:1 insert/vector molar ratio.

    Article Title: Mucosal and systemic immune responses elicited by recombinant Lactococcus lactis expressing a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis
    Article Snippet: .. The synthesized fragment (SECF1S1) was digested with Kpn I and Xba I restriction endonucleases (Thermo Fisher Scientific) and was inserted into the corresponding sites of the expression vector pNZ8149, giving rise to pNZ-SECF1S1. ..

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  • 99
    Thermo Fisher kpn i
    Identification of the recombinant plasmid <t>pNZ8149-SECF1S1</t> by PCR and restriction enzyme analysis (a) PCR amplification of F1S1gene. Lane A: DNA ladder; Lanes B and C: PCR products of the F1S1 gene amplification D: No template control. (b) Restriction analysis of the recombinant plasmid pNZ8149-SECF1S1. Lane A: DNA ladder; Lane B: pNZ8149-SECF1S1 double-digested with <t>Kpn</t> I and Xba I; C: pNZ8149-SECF1S1 digested with Kpn I.
    Kpn I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 330 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/kpn i/product/Thermo Fisher
    Average 99 stars, based on 330 article reviews
    Price from $9.99 to $1999.99
    kpn i - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    90
    Thermo Fisher restriction endonucleases kpn i
    Gel electrophoresis of recombinant plasmid following <t>Kpn</t> I and Xho I double enzymatic digestion. PSMA, prostate-specific membrane antigen.
    Restriction Endonucleases Kpn I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction endonucleases kpn i/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction endonucleases kpn i - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    Image Search Results


    Identification of the recombinant plasmid pNZ8149-SECF1S1 by PCR and restriction enzyme analysis (a) PCR amplification of F1S1gene. Lane A: DNA ladder; Lanes B and C: PCR products of the F1S1 gene amplification D: No template control. (b) Restriction analysis of the recombinant plasmid pNZ8149-SECF1S1. Lane A: DNA ladder; Lane B: pNZ8149-SECF1S1 double-digested with Kpn I and Xba I; C: pNZ8149-SECF1S1 digested with Kpn I.

    Journal: Microbial Pathogenesis

    Article Title: Mucosal and systemic immune responses elicited by recombinant Lactococcus lactis expressing a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis

    doi: 10.1016/j.micpath.2018.05.008

    Figure Lengend Snippet: Identification of the recombinant plasmid pNZ8149-SECF1S1 by PCR and restriction enzyme analysis (a) PCR amplification of F1S1gene. Lane A: DNA ladder; Lanes B and C: PCR products of the F1S1 gene amplification D: No template control. (b) Restriction analysis of the recombinant plasmid pNZ8149-SECF1S1. Lane A: DNA ladder; Lane B: pNZ8149-SECF1S1 double-digested with Kpn I and Xba I; C: pNZ8149-SECF1S1 digested with Kpn I.

    Article Snippet: The synthesized fragment (SECF1S1) was digested with Kpn I and Xba I restriction endonucleases (Thermo Fisher Scientific) and was inserted into the corresponding sites of the expression vector pNZ8149, giving rise to pNZ-SECF1S1.

    Techniques: Recombinant, Plasmid Preparation, Polymerase Chain Reaction, Amplification

    Cloning and sequence alignment of D. melanogaster HDs for bacterial overexpression. Vector map (A) and sequence map (B) of HisSUMO-HD constructs. Each of the HDs for FTZ, ABD-A, ABD-B, ANTP, and UBX were cloned into a T7-inducible HisSUMO vector between Kpn I and Xba I restriction sites. (C) Sequence alignment of overexpressed D. melanogaster ]. Only the five HDs studied in this paper are shown. Highly conserved residues within similar physicochemical properties in all 107 HDs analysed are shown (red – positive, blue – negative, yellow – polar, green – hydrophobic). Absolutely conserved residues and residues with greater than 70% similarity are labelled with a star or cross, respectively.

    Journal: Protein expression and purification

    Article Title: Rapid and Efficient Purification of Drosophila Homeodomain Transcription Factors for Biophysical Characterization

    doi: 10.1016/j.pep.2019.02.001

    Figure Lengend Snippet: Cloning and sequence alignment of D. melanogaster HDs for bacterial overexpression. Vector map (A) and sequence map (B) of HisSUMO-HD constructs. Each of the HDs for FTZ, ABD-A, ABD-B, ANTP, and UBX were cloned into a T7-inducible HisSUMO vector between Kpn I and Xba I restriction sites. (C) Sequence alignment of overexpressed D. melanogaster ]. Only the five HDs studied in this paper are shown. Highly conserved residues within similar physicochemical properties in all 107 HDs analysed are shown (red – positive, blue – negative, yellow – polar, green – hydrophobic). Absolutely conserved residues and residues with greater than 70% similarity are labelled with a star or cross, respectively.

    Article Snippet: The HD domains of UBX ( , residues 387–446), AbdA ( , residues 398–457), and AbdB ( , residues 295–354) were PCR amplified using KOD polymerase (Novagen), digested with Kpn I and XbaI, and ligated into the compatible restriction sites of the modified HisSUMO vector using T4 DNA ligase (Thermo-Fisher).

    Techniques: Clone Assay, Sequencing, Over Expression, Plasmid Preparation, Construct

    Gel electrophoresis of recombinant plasmid following Kpn I and Xho I double enzymatic digestion. PSMA, prostate-specific membrane antigen.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Expression, purification and identification of an immunogenic fragment in the ectodomain of prostate-specific membrane antigen

    doi: 10.3892/etm.2016.3005

    Figure Lengend Snippet: Gel electrophoresis of recombinant plasmid following Kpn I and Xho I double enzymatic digestion. PSMA, prostate-specific membrane antigen.

    Article Snippet: Reagents Reagents used were sourced as follows: Restriction endonucleases Kpn I and Xho I, RPMI-1650, fetal bovine serum (Thermo Fisher Scientific, Inc., Waltham, MA, USA); T4 ligase, isopropyl-β-D-thiogalactopyranoside (IPTG), a 10 kb DNA ladder (cat. no. B600024l; Sangon Biotech Co., Ltd., Shanghai, China) a protein marker (cat. no. SM0431; Thermo Fisher Scientific, Inc.), an EZNA plasmid extraction kit (cat. no. D6943; Omega Bio-Tek Inc., Norcross, GA, USA); a plasmid purification kit (Merck Sharpe & Dohme, Shanghai, China); acrylamide and methylene bis-acrylamide (Genview Scientific Inc., El Monte, CA, USA); a 3,000-unit dialysis bag, sodium dodecyl sulfate (SDS) and Tris base (Sino-American Biotechnology Co., Ltd., Shanghai, China); anti-PSMA (YPSMA-1) mouse monoclonal antibody (cat. no. ab19071, Abcam, Shanghai, China); and Freund's complete adjuvant and fluorescein isothiocyanate (FITC)-labeled goat anti-mouse polyclonal immunoglobulin G (cat. no. F9006; Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: Nucleic Acid Electrophoresis, Recombinant, Plasmid Preparation