pmd19 vector  (TaKaRa)

 
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    Name:
    T Vector pMD 19
    Description:
    pMD20 and pMD19 Simple T Vectors are linearized PCR cloning vectors cleaved by EcoRV with single 3 terminal thymidine residues dT at both ends These T overhangs at the cloning site improve the efficiency of ligation of PCR products that contain dA overhangs The inclusion of lacZ in these vectors allows blue white screening for the detection of successful ligations
    Catalog Number:
    3271
    Price:
    None
    Size:
    1 ug
    Category:
    T Vectors pMD20 and pMD19 Cloning vectors Linkers primers and cloning vectors Cloning
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    Structured Review

    TaKaRa pmd19 vector
    pMD20 and pMD19 Simple T Vectors are linearized PCR cloning vectors cleaved by EcoRV with single 3 terminal thymidine residues dT at both ends These T overhangs at the cloning site improve the efficiency of ligation of PCR products that contain dA overhangs The inclusion of lacZ in these vectors allows blue white screening for the detection of successful ligations
    https://www.bioz.com/result/pmd19 vector/product/TaKaRa
    Average 99 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    pmd19 vector - by Bioz Stars, 2020-08
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane
    Article Snippet: .. The PCR amplification procedure was: 94 °C initial denaturation for 4 min; 94 °C denaturation for 30 s, 52 °C annealed for 30 s, 72 °C extension for 30 s, cycled 35 times; 72 °C re-extension for 10 min. After purification by gel extraction, all PCR amplification products (507 bp) were built onto a pMD19-T Cloning Vector (Takara, China) and transformed into Escheria coli DH5α. ..

    Article Title: Establishment of a Successive Markerless Mutation System in Haemophilus parasuis through Natural Transformation
    Article Snippet: .. The three PCR fragments were mixed with pMD19-T vector (Takara, Japan) and ligated using ClonExpress MultiS One Step Cloning Kit (Vazyme, China) according to the manufacturer’s protocol. .. The resulting products were transformed into E . coli DH5α and transformants were selected on LB agar containing 50 μg/mL Kan. PCRs were performed on selected colonies to confirm the presence of inserts.

    Article Title: Establishment of a Successive Markerless Mutation System in Haemophilus parasuis through Natural Transformation
    Article Snippet: .. The two PCR fragments were ligated with pMD19-T vector using ClonExpress MultiS One Step Cloning Kit to form pMDP. ..

    Amplification:

    Article Title: Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane
    Article Snippet: .. The PCR amplification procedure was: 94 °C initial denaturation for 4 min; 94 °C denaturation for 30 s, 52 °C annealed for 30 s, 72 °C extension for 30 s, cycled 35 times; 72 °C re-extension for 10 min. After purification by gel extraction, all PCR amplification products (507 bp) were built onto a pMD19-T Cloning Vector (Takara, China) and transformed into Escheria coli DH5α. ..

    Synthesized:

    Article Title: Recombination and identification of human alpha B-crystallin
    Article Snippet: .. The synthesized target DNA fragment was ligated into the PMD19-T vector in order to construct the recombinant plasmid PMD19-T-αB-crystallin, which was then digested by restriction endonuclease Ecor l and XhoI. .. Gel electrophoresis revealed that the cleavage of Ecor l and XhoI generated the target fragment, which was consistent with the expected fragment size of 545 bp ( ).

    Article Title: Recombination and identification of human alpha B-crystallin
    Article Snippet: .. The synthesized gene fragment was then inserted into the PMD19-T vector at restriction enzyme sites EcoRI and XhoI. .. The products were transformed into E. coli BL21 (DE3) pLysS.

    Construct:

    Article Title: Recombination and identification of human alpha B-crystallin
    Article Snippet: .. The synthesized target DNA fragment was ligated into the PMD19-T vector in order to construct the recombinant plasmid PMD19-T-αB-crystallin, which was then digested by restriction endonuclease Ecor l and XhoI. .. Gel electrophoresis revealed that the cleavage of Ecor l and XhoI generated the target fragment, which was consistent with the expected fragment size of 545 bp ( ).

    Purification:

    Article Title: Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane
    Article Snippet: .. The PCR amplification procedure was: 94 °C initial denaturation for 4 min; 94 °C denaturation for 30 s, 52 °C annealed for 30 s, 72 °C extension for 30 s, cycled 35 times; 72 °C re-extension for 10 min. After purification by gel extraction, all PCR amplification products (507 bp) were built onto a pMD19-T Cloning Vector (Takara, China) and transformed into Escheria coli DH5α. ..

    Sequencing:

    Article Title: CYP1A1 Relieves Lipopolysaccharide-Induced Inflammatory Responses in Bovine Mammary Epithelial Cells
    Article Snippet: .. The product with added polyA was further collected using a StarPrep Gel Extraction Kit (GenStar BioSolutions) and connected to a pMD19-T vector (TakaRa) by using T4 DNA Ligase (TakaRa) at 4°C for 12 h. Finally, the plasmids from the transformants of Trans5 α containing the sequence for the molecular cloning of full-length bovine CYP1A1 gene were collected using a TIANprep Rapid Mini Plasmid Kit (TIANGEN, Shanghai, China) and sequenced by BIG Tech (Shenzhen, China) to verify the correct DNA sequence. .. The plasmid of the pMD19-T vector connected with the correct sequence was digested by restriction enzymes Nhel and Notl (New England Biolabs, Ipswich, MA).

    Molecular Cloning:

    Article Title: CYP1A1 Relieves Lipopolysaccharide-Induced Inflammatory Responses in Bovine Mammary Epithelial Cells
    Article Snippet: .. The product with added polyA was further collected using a StarPrep Gel Extraction Kit (GenStar BioSolutions) and connected to a pMD19-T vector (TakaRa) by using T4 DNA Ligase (TakaRa) at 4°C for 12 h. Finally, the plasmids from the transformants of Trans5 α containing the sequence for the molecular cloning of full-length bovine CYP1A1 gene were collected using a TIANprep Rapid Mini Plasmid Kit (TIANGEN, Shanghai, China) and sequenced by BIG Tech (Shenzhen, China) to verify the correct DNA sequence. .. The plasmid of the pMD19-T vector connected with the correct sequence was digested by restriction enzymes Nhel and Notl (New England Biolabs, Ipswich, MA).

    Polymerase Chain Reaction:

    Article Title: Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane
    Article Snippet: .. The PCR amplification procedure was: 94 °C initial denaturation for 4 min; 94 °C denaturation for 30 s, 52 °C annealed for 30 s, 72 °C extension for 30 s, cycled 35 times; 72 °C re-extension for 10 min. After purification by gel extraction, all PCR amplification products (507 bp) were built onto a pMD19-T Cloning Vector (Takara, China) and transformed into Escheria coli DH5α. ..

    Article Title: Establishment of a Successive Markerless Mutation System in Haemophilus parasuis through Natural Transformation
    Article Snippet: .. The three PCR fragments were mixed with pMD19-T vector (Takara, Japan) and ligated using ClonExpress MultiS One Step Cloning Kit (Vazyme, China) according to the manufacturer’s protocol. .. The resulting products were transformed into E . coli DH5α and transformants were selected on LB agar containing 50 μg/mL Kan. PCRs were performed on selected colonies to confirm the presence of inserts.

    Article Title: Establishment of a Successive Markerless Mutation System in Haemophilus parasuis through Natural Transformation
    Article Snippet: .. The two PCR fragments were ligated with pMD19-T vector using ClonExpress MultiS One Step Cloning Kit to form pMDP. ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Cell Signaling Pathway in 12-O-Tetradecanoylphorbol-13-acetate-Induced LCN2 Gene Transcription in Esophageal Squamous Cell Carcinoma
    Article Snippet: .. The pMD19-T Simple vector, SYBR Premix Ex-Taq™, and SYBR Primescript RT-PCR Kit were purchased from TaKaRa (Dalian, China). .. Expression Vector Construction and Stable Transfection The full-length cDNAs for MISP, KLF10, KLF15, PPP1R18, and RXRβ were amplified by RT-PCR.

    Gel Extraction:

    Article Title: Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane
    Article Snippet: .. The PCR amplification procedure was: 94 °C initial denaturation for 4 min; 94 °C denaturation for 30 s, 52 °C annealed for 30 s, 72 °C extension for 30 s, cycled 35 times; 72 °C re-extension for 10 min. After purification by gel extraction, all PCR amplification products (507 bp) were built onto a pMD19-T Cloning Vector (Takara, China) and transformed into Escheria coli DH5α. ..

    Article Title: CYP1A1 Relieves Lipopolysaccharide-Induced Inflammatory Responses in Bovine Mammary Epithelial Cells
    Article Snippet: .. The product with added polyA was further collected using a StarPrep Gel Extraction Kit (GenStar BioSolutions) and connected to a pMD19-T vector (TakaRa) by using T4 DNA Ligase (TakaRa) at 4°C for 12 h. Finally, the plasmids from the transformants of Trans5 α containing the sequence for the molecular cloning of full-length bovine CYP1A1 gene were collected using a TIANprep Rapid Mini Plasmid Kit (TIANGEN, Shanghai, China) and sequenced by BIG Tech (Shenzhen, China) to verify the correct DNA sequence. .. The plasmid of the pMD19-T vector connected with the correct sequence was digested by restriction enzymes Nhel and Notl (New England Biolabs, Ipswich, MA).

    Transformation Assay:

    Article Title: Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane
    Article Snippet: .. The PCR amplification procedure was: 94 °C initial denaturation for 4 min; 94 °C denaturation for 30 s, 52 °C annealed for 30 s, 72 °C extension for 30 s, cycled 35 times; 72 °C re-extension for 10 min. After purification by gel extraction, all PCR amplification products (507 bp) were built onto a pMD19-T Cloning Vector (Takara, China) and transformed into Escheria coli DH5α. ..

    Recombinant:

    Article Title: Recombination and identification of human alpha B-crystallin
    Article Snippet: .. The synthesized target DNA fragment was ligated into the PMD19-T vector in order to construct the recombinant plasmid PMD19-T-αB-crystallin, which was then digested by restriction endonuclease Ecor l and XhoI. .. Gel electrophoresis revealed that the cleavage of Ecor l and XhoI generated the target fragment, which was consistent with the expected fragment size of 545 bp ( ).

    Plasmid Preparation:

    Article Title: Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane
    Article Snippet: .. The PCR amplification procedure was: 94 °C initial denaturation for 4 min; 94 °C denaturation for 30 s, 52 °C annealed for 30 s, 72 °C extension for 30 s, cycled 35 times; 72 °C re-extension for 10 min. After purification by gel extraction, all PCR amplification products (507 bp) were built onto a pMD19-T Cloning Vector (Takara, China) and transformed into Escheria coli DH5α. ..

    Article Title: Recombination and identification of human alpha B-crystallin
    Article Snippet: .. The synthesized target DNA fragment was ligated into the PMD19-T vector in order to construct the recombinant plasmid PMD19-T-αB-crystallin, which was then digested by restriction endonuclease Ecor l and XhoI. .. Gel electrophoresis revealed that the cleavage of Ecor l and XhoI generated the target fragment, which was consistent with the expected fragment size of 545 bp ( ).

    Article Title: CYP1A1 Relieves Lipopolysaccharide-Induced Inflammatory Responses in Bovine Mammary Epithelial Cells
    Article Snippet: .. The dual-enzyme digestion of the plasmid of pMD19-T–CYP1A1 cDNA confirmed that CYP1A1 cDNA was accurately connected into the pMD19-T vector ( ). .. To check the integrity and accuracy of the CYP1A1 cDNA, the cloned full-length bovine CYP1A1 cDNA was sequenced and blasted with the complete DNA sequences of encoding CYP1A1 protein in the NCBI database.

    Article Title: CYP1A1 Relieves Lipopolysaccharide-Induced Inflammatory Responses in Bovine Mammary Epithelial Cells
    Article Snippet: .. The product with added polyA was further collected using a StarPrep Gel Extraction Kit (GenStar BioSolutions) and connected to a pMD19-T vector (TakaRa) by using T4 DNA Ligase (TakaRa) at 4°C for 12 h. Finally, the plasmids from the transformants of Trans5 α containing the sequence for the molecular cloning of full-length bovine CYP1A1 gene were collected using a TIANprep Rapid Mini Plasmid Kit (TIANGEN, Shanghai, China) and sequenced by BIG Tech (Shenzhen, China) to verify the correct DNA sequence. .. The plasmid of the pMD19-T vector connected with the correct sequence was digested by restriction enzymes Nhel and Notl (New England Biolabs, Ipswich, MA).

    Article Title: Recombination and identification of human alpha B-crystallin
    Article Snippet: .. The synthesized gene fragment was then inserted into the PMD19-T vector at restriction enzyme sites EcoRI and XhoI. .. The products were transformed into E. coli BL21 (DE3) pLysS.

    Article Title: Establishment of a Successive Markerless Mutation System in Haemophilus parasuis through Natural Transformation
    Article Snippet: .. The three PCR fragments were mixed with pMD19-T vector (Takara, Japan) and ligated using ClonExpress MultiS One Step Cloning Kit (Vazyme, China) according to the manufacturer’s protocol. .. The resulting products were transformed into E . coli DH5α and transformants were selected on LB agar containing 50 μg/mL Kan. PCRs were performed on selected colonies to confirm the presence of inserts.

    Article Title: Establishment of a Successive Markerless Mutation System in Haemophilus parasuis through Natural Transformation
    Article Snippet: .. The two PCR fragments were ligated with pMD19-T vector using ClonExpress MultiS One Step Cloning Kit to form pMDP. ..

    Article Title: Cell Signaling Pathway in 12-O-Tetradecanoylphorbol-13-acetate-Induced LCN2 Gene Transcription in Esophageal Squamous Cell Carcinoma
    Article Snippet: .. The pMD19-T Simple vector, SYBR Premix Ex-Taq™, and SYBR Primescript RT-PCR Kit were purchased from TaKaRa (Dalian, China). .. Expression Vector Construction and Stable Transfection The full-length cDNAs for MISP, KLF10, KLF15, PPP1R18, and RXRβ were amplified by RT-PCR.

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  • 99
    TaKaRa pmd19 t vector
    Construction of CYP1A1 overexpression plasmids and verification of overexpression efficiency. (a) Cloned full-length CYP1A1 cDNA from MAC-T cells was analyzed using PCR. (b) The dual-enzyme digestion of the plasmid of <t>pMD19-T–CYP1A1</t> cDNA confirmed that CYP1A1 cDNA was accurately connected into the pMD19-T vector. (c) The efficient connection of cloned CYP1A1 cDNAs to the target plasmid was confirmed using plasmid PCR. Control: empty vector. 1, 2, and 3: target vector connected to CYP1A1 from three bacterial colonies. (d) Overexpression efficiency of CYP1A1 was verified using RT-qPCR and Western blot. GAPDH was used as an internal control. Data were expressed as the mean ± SD. ∗ p
    Pmd19 T Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1088 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmd19 t vector/product/TaKaRa
    Average 99 stars, based on 1088 article reviews
    Price from $9.99 to $1999.99
    pmd19 t vector - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

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    Construction of CYP1A1 overexpression plasmids and verification of overexpression efficiency. (a) Cloned full-length CYP1A1 cDNA from MAC-T cells was analyzed using PCR. (b) The dual-enzyme digestion of the plasmid of pMD19-T–CYP1A1 cDNA confirmed that CYP1A1 cDNA was accurately connected into the pMD19-T vector. (c) The efficient connection of cloned CYP1A1 cDNAs to the target plasmid was confirmed using plasmid PCR. Control: empty vector. 1, 2, and 3: target vector connected to CYP1A1 from three bacterial colonies. (d) Overexpression efficiency of CYP1A1 was verified using RT-qPCR and Western blot. GAPDH was used as an internal control. Data were expressed as the mean ± SD. ∗ p

    Journal: Mediators of Inflammation

    Article Title: CYP1A1 Relieves Lipopolysaccharide-Induced Inflammatory Responses in Bovine Mammary Epithelial Cells

    doi: 10.1155/2018/4093285

    Figure Lengend Snippet: Construction of CYP1A1 overexpression plasmids and verification of overexpression efficiency. (a) Cloned full-length CYP1A1 cDNA from MAC-T cells was analyzed using PCR. (b) The dual-enzyme digestion of the plasmid of pMD19-T–CYP1A1 cDNA confirmed that CYP1A1 cDNA was accurately connected into the pMD19-T vector. (c) The efficient connection of cloned CYP1A1 cDNAs to the target plasmid was confirmed using plasmid PCR. Control: empty vector. 1, 2, and 3: target vector connected to CYP1A1 from three bacterial colonies. (d) Overexpression efficiency of CYP1A1 was verified using RT-qPCR and Western blot. GAPDH was used as an internal control. Data were expressed as the mean ± SD. ∗ p

    Article Snippet: The dual-enzyme digestion of the plasmid of pMD19-T–CYP1A1 cDNA confirmed that CYP1A1 cDNA was accurately connected into the pMD19-T vector ( ).

    Techniques: Over Expression, Polymerase Chain Reaction, Plasmid Preparation, Clone Assay, Quantitative RT-PCR, Western Blot

    Scheme for the construction of plasmid pMDHK. Primer pairs P1/P2 and P3/P4 were used to amplify the upstream and downstream regions of the htrA gene respectively, each with 20-bp end sequences identical to the kan fragment (using primer P2 and P3) and 20-bp end sequences identical to vector pMD19-T (using primer P1 and P4). Primers P5/P6 were used to amplify the kan fragment from the plasmid pKD4. The resulting three PCR products and the cloning vector pMD19-T were joined by In-Fusion cloning. Two USSs required for efficient transformation were introduced into the plasmid pMDHK by using primer P1 and P4.

    Journal: PLoS ONE

    Article Title: Establishment of a Successive Markerless Mutation System in Haemophilus parasuis through Natural Transformation

    doi: 10.1371/journal.pone.0127393

    Figure Lengend Snippet: Scheme for the construction of plasmid pMDHK. Primer pairs P1/P2 and P3/P4 were used to amplify the upstream and downstream regions of the htrA gene respectively, each with 20-bp end sequences identical to the kan fragment (using primer P2 and P3) and 20-bp end sequences identical to vector pMD19-T (using primer P1 and P4). Primers P5/P6 were used to amplify the kan fragment from the plasmid pKD4. The resulting three PCR products and the cloning vector pMD19-T were joined by In-Fusion cloning. Two USSs required for efficient transformation were introduced into the plasmid pMDHK by using primer P1 and P4.

    Article Snippet: The three PCR fragments were mixed with pMD19-T vector (Takara, Japan) and ligated using ClonExpress MultiS One Step Cloning Kit (Vazyme, China) according to the manufacturer’s protocol.

    Techniques: Plasmid Preparation, Polymerase Chain Reaction, Clone Assay, Transformation Assay

    Gel electrophoresis following Ecor l and XhoI double enzymatic digestion A: Recombinant plasmid PMD19-T-αB-crystallin; B: Recombinant plasmid pET28a-αB-crystallin. Lane 1: Marker; Lane 2: Target gene fragment.

    Journal: International Journal of Ophthalmology

    Article Title: Recombination and identification of human alpha B-crystallin

    doi: 10.18240/ijo.2018.12.06

    Figure Lengend Snippet: Gel electrophoresis following Ecor l and XhoI double enzymatic digestion A: Recombinant plasmid PMD19-T-αB-crystallin; B: Recombinant plasmid pET28a-αB-crystallin. Lane 1: Marker; Lane 2: Target gene fragment.

    Article Snippet: The synthesized target DNA fragment was ligated into the PMD19-T vector in order to construct the recombinant plasmid PMD19-T-αB-crystallin, which was then digested by restriction endonuclease Ecor l and XhoI.

    Techniques: Nucleic Acid Electrophoresis, Recombinant, Plasmid Preparation, Marker