vector pgem t  (Promega)

 
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    Name:
    pGEM T Vector Systems
    Description:
    T vector for simplified cloning of PCR products
    Catalog Number:
    a3600
    Price:
    None
    Category:
    Nucleic Acid Extraction Analysis PCR PCR Cloning
    Buy from Supplier


    Structured Review

    Promega vector pgem t
    Expression of recombinant MAA2 in E. coli . (A) E. coli cells containing the maa2 gene ligated into vector <t>pGEM-T</t> in both orientations with respect to the lacZ promoter were grown in the presence and absence of IPTG, lysed, electrophoresed by SDS-PAGE on 7.5% (wt/vol) gels, transferred to nitrocellulose, and stained with MAA2-specific MAb 7a. Lanes contain M. arthritidis 158p10p9 whole-cell lysate (lane 1), E. coli containing the cloned insert in the forward orientation with respect to the lacZ promoter, in the absence (lane 2) and presence (lane 3) of IPTG, E. coli containing the cloned insert in the reverse orientation with respect to the lacZ promoter, in the absence (lane 4) and presence (lane 5) of IPTG, and untransformed E. coli in the absence (lane 6) and presence (lane 7) of IPTG. (B) E. coli cells containing the maa2 gene from M. arthritidis 158p10p9 clonal variants in which expression of MAA2 was switched ON or OFF were grown in the presence or absence of IPTG, lysed, electrophoresed and immunoblotted as described above. Lanes contain E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the reverse orientation with respect to the lacZ promoter, in the absence (lane 1) and presence (lane 2) of IPTG, E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 3) and presence (lane 4) of IPTG, E. coli in which maa2 from an ON variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 5) and presence (lane 6) of IPTG, and a whole-cell lysate of M. arthritidis strain 158p10p9 (lane 7).
    T vector for simplified cloning of PCR products
    https://www.bioz.com/result/vector pgem t/product/Promega
    Average 99 stars, based on 53 article reviews
    Price from $9.99 to $1999.99
    vector pgem t - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "Molecular Characterization of Mycoplasma arthritidis Variable Surface Protein MAA2"

    Article Title: Molecular Characterization of Mycoplasma arthritidis Variable Surface Protein MAA2

    Journal: Infection and Immunity

    doi:

    Expression of recombinant MAA2 in E. coli . (A) E. coli cells containing the maa2 gene ligated into vector pGEM-T in both orientations with respect to the lacZ promoter were grown in the presence and absence of IPTG, lysed, electrophoresed by SDS-PAGE on 7.5% (wt/vol) gels, transferred to nitrocellulose, and stained with MAA2-specific MAb 7a. Lanes contain M. arthritidis 158p10p9 whole-cell lysate (lane 1), E. coli containing the cloned insert in the forward orientation with respect to the lacZ promoter, in the absence (lane 2) and presence (lane 3) of IPTG, E. coli containing the cloned insert in the reverse orientation with respect to the lacZ promoter, in the absence (lane 4) and presence (lane 5) of IPTG, and untransformed E. coli in the absence (lane 6) and presence (lane 7) of IPTG. (B) E. coli cells containing the maa2 gene from M. arthritidis 158p10p9 clonal variants in which expression of MAA2 was switched ON or OFF were grown in the presence or absence of IPTG, lysed, electrophoresed and immunoblotted as described above. Lanes contain E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the reverse orientation with respect to the lacZ promoter, in the absence (lane 1) and presence (lane 2) of IPTG, E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 3) and presence (lane 4) of IPTG, E. coli in which maa2 from an ON variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 5) and presence (lane 6) of IPTG, and a whole-cell lysate of M. arthritidis strain 158p10p9 (lane 7).
    Figure Legend Snippet: Expression of recombinant MAA2 in E. coli . (A) E. coli cells containing the maa2 gene ligated into vector pGEM-T in both orientations with respect to the lacZ promoter were grown in the presence and absence of IPTG, lysed, electrophoresed by SDS-PAGE on 7.5% (wt/vol) gels, transferred to nitrocellulose, and stained with MAA2-specific MAb 7a. Lanes contain M. arthritidis 158p10p9 whole-cell lysate (lane 1), E. coli containing the cloned insert in the forward orientation with respect to the lacZ promoter, in the absence (lane 2) and presence (lane 3) of IPTG, E. coli containing the cloned insert in the reverse orientation with respect to the lacZ promoter, in the absence (lane 4) and presence (lane 5) of IPTG, and untransformed E. coli in the absence (lane 6) and presence (lane 7) of IPTG. (B) E. coli cells containing the maa2 gene from M. arthritidis 158p10p9 clonal variants in which expression of MAA2 was switched ON or OFF were grown in the presence or absence of IPTG, lysed, electrophoresed and immunoblotted as described above. Lanes contain E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the reverse orientation with respect to the lacZ promoter, in the absence (lane 1) and presence (lane 2) of IPTG, E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 3) and presence (lane 4) of IPTG, E. coli in which maa2 from an ON variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 5) and presence (lane 6) of IPTG, and a whole-cell lysate of M. arthritidis strain 158p10p9 (lane 7).

    Techniques Used: Expressing, Recombinant, Plasmid Preparation, SDS Page, Staining, Clone Assay, Variant Assay

    Related Articles

    Clone Assay:

    Article Title: Chromosome-Based Genetic Complementation System for Xylella fastidiosa ▿
    Article Snippet: .. The resulting 1.6-kb fragment was then cloned into the pGEM-T vector (Promega), creating plasmid pAX1. .. PCR was also used to generate DNA fragments carrying the multiple cloning sites and the four antibiotic resistance cassettes that were inserted into pAX1.

    Article Title: Cloning and molecular characterization of Δ12-fatty acid desaturase gene from Mortierella isabellina
    Article Snippet: .. PCR product about 1.2 kb long was gel-purified by electrophoresis and cloned into pGEM-T vector. .. Positive clones were selected on LB plate by color reaction.

    Article Title: Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale
    Article Snippet: .. In the case of mixed infections (presence of double peaks in the analyzed pherograms from the sequencing of the original PCR amplicon), purified PCR products were ligated into pGEM-T easy cloning vector following the manufacturer's instructions (pGEM-T clone kit; Promega, Madison, WI) and transformed into Escherichia coli DH5α competent cells by heat shock. .. Positive colonies were selected by blue/white screening followed by colony PCR using M13F/R primers under the following conditions: 5 min at 95°C and 35 cycles of 60 s at 95°C, 60 s at 51°C, and 1 min and 20 s at 72°C and a final extension of 5 min at 72°C.

    Article Title: Motifs within the CA-repeat-rich region of Surfactant Protein B (SFTPB) intron 4 differentially affect mRNA splicing
    Article Snippet: .. To improve the specificity of amplification in order to generate different length variants, the following touchdown PCR program ( ) was used: 95°C for 2 min, 5 cycles of 95°C for 30 s, 70°C for 30 s and 68°C for 30 s, then 5 cycles of 95°C for 30 s, 68°C for 30 s and 68°C for 30 s, and then 5 cycles of 95°C for 30 s, 66°C for 30 s and 68°C for 30 s, followed by 25 cycles of 94°C for 30 s, 64°C for 30 s and 68°C for 30 s and a final extension step at 68°C for 5 min. All PCR fragments, ranging from 96 to 437 were separated on agarose gel, purified and cloned into a pGEM-T vector (Promega, WI, USA). .. The recombinant plasmid DNAs were digested, purified and sub-cloned into the EcoR I and Not I sites of pcDNA 3.1 minigene Del-m ( , inset).

    Article Title: Two RNA editing sites with cis-acting elements of moderate sequence identity are recognized by an identical site-recognition protein in tobacco chloroplasts
    Article Snippet: .. The amplified fragments were cloned into a pGEM-T vector using the pGEM-T Vector System (Promega). .. From these cloned plasmids, RNA substrates for in vitro editing and UV-crosslinking were prepared as previously described ( ) with slight modifications.

    Article Title: A gene encoding a protein modified by the phytohormone indoleacetic acid
    Article Snippet: .. The iap1 cDNA was cloned into a pGEM-T vector and was used for coupled in vitro transcription and translation using the TnT Quick Coupled System (Promega) with [35 S]methionine. .. GeneRacer Kit (Invitrogen) was used for full-length, RNA ligase-mediated rapid amplification of 5′ ends using RNA isolated 24 days after flowering.

    Amplification:

    Article Title: Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale
    Article Snippet: .. In the case of mixed infections (presence of double peaks in the analyzed pherograms from the sequencing of the original PCR amplicon), purified PCR products were ligated into pGEM-T easy cloning vector following the manufacturer's instructions (pGEM-T clone kit; Promega, Madison, WI) and transformed into Escherichia coli DH5α competent cells by heat shock. .. Positive colonies were selected by blue/white screening followed by colony PCR using M13F/R primers under the following conditions: 5 min at 95°C and 35 cycles of 60 s at 95°C, 60 s at 51°C, and 1 min and 20 s at 72°C and a final extension of 5 min at 72°C.

    Article Title: Motifs within the CA-repeat-rich region of Surfactant Protein B (SFTPB) intron 4 differentially affect mRNA splicing
    Article Snippet: .. To improve the specificity of amplification in order to generate different length variants, the following touchdown PCR program ( ) was used: 95°C for 2 min, 5 cycles of 95°C for 30 s, 70°C for 30 s and 68°C for 30 s, then 5 cycles of 95°C for 30 s, 68°C for 30 s and 68°C for 30 s, and then 5 cycles of 95°C for 30 s, 66°C for 30 s and 68°C for 30 s, followed by 25 cycles of 94°C for 30 s, 64°C for 30 s and 68°C for 30 s and a final extension step at 68°C for 5 min. All PCR fragments, ranging from 96 to 437 were separated on agarose gel, purified and cloned into a pGEM-T vector (Promega, WI, USA). .. The recombinant plasmid DNAs were digested, purified and sub-cloned into the EcoR I and Not I sites of pcDNA 3.1 minigene Del-m ( , inset).

    Article Title: Two RNA editing sites with cis-acting elements of moderate sequence identity are recognized by an identical site-recognition protein in tobacco chloroplasts
    Article Snippet: .. The amplified fragments were cloned into a pGEM-T vector using the pGEM-T Vector System (Promega). .. From these cloned plasmids, RNA substrates for in vitro editing and UV-crosslinking were prepared as previously described ( ) with slight modifications.

    Agarose Gel Electrophoresis:

    Article Title: Motifs within the CA-repeat-rich region of Surfactant Protein B (SFTPB) intron 4 differentially affect mRNA splicing
    Article Snippet: .. To improve the specificity of amplification in order to generate different length variants, the following touchdown PCR program ( ) was used: 95°C for 2 min, 5 cycles of 95°C for 30 s, 70°C for 30 s and 68°C for 30 s, then 5 cycles of 95°C for 30 s, 68°C for 30 s and 68°C for 30 s, and then 5 cycles of 95°C for 30 s, 66°C for 30 s and 68°C for 30 s, followed by 25 cycles of 94°C for 30 s, 64°C for 30 s and 68°C for 30 s and a final extension step at 68°C for 5 min. All PCR fragments, ranging from 96 to 437 were separated on agarose gel, purified and cloned into a pGEM-T vector (Promega, WI, USA). .. The recombinant plasmid DNAs were digested, purified and sub-cloned into the EcoR I and Not I sites of pcDNA 3.1 minigene Del-m ( , inset).

    In Vitro:

    Article Title: A gene encoding a protein modified by the phytohormone indoleacetic acid
    Article Snippet: .. The iap1 cDNA was cloned into a pGEM-T vector and was used for coupled in vitro transcription and translation using the TnT Quick Coupled System (Promega) with [35 S]methionine. .. GeneRacer Kit (Invitrogen) was used for full-length, RNA ligase-mediated rapid amplification of 5′ ends using RNA isolated 24 days after flowering.

    Purification:

    Article Title: Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale
    Article Snippet: .. In the case of mixed infections (presence of double peaks in the analyzed pherograms from the sequencing of the original PCR amplicon), purified PCR products were ligated into pGEM-T easy cloning vector following the manufacturer's instructions (pGEM-T clone kit; Promega, Madison, WI) and transformed into Escherichia coli DH5α competent cells by heat shock. .. Positive colonies were selected by blue/white screening followed by colony PCR using M13F/R primers under the following conditions: 5 min at 95°C and 35 cycles of 60 s at 95°C, 60 s at 51°C, and 1 min and 20 s at 72°C and a final extension of 5 min at 72°C.

    Article Title: Motifs within the CA-repeat-rich region of Surfactant Protein B (SFTPB) intron 4 differentially affect mRNA splicing
    Article Snippet: .. To improve the specificity of amplification in order to generate different length variants, the following touchdown PCR program ( ) was used: 95°C for 2 min, 5 cycles of 95°C for 30 s, 70°C for 30 s and 68°C for 30 s, then 5 cycles of 95°C for 30 s, 68°C for 30 s and 68°C for 30 s, and then 5 cycles of 95°C for 30 s, 66°C for 30 s and 68°C for 30 s, followed by 25 cycles of 94°C for 30 s, 64°C for 30 s and 68°C for 30 s and a final extension step at 68°C for 5 min. All PCR fragments, ranging from 96 to 437 were separated on agarose gel, purified and cloned into a pGEM-T vector (Promega, WI, USA). .. The recombinant plasmid DNAs were digested, purified and sub-cloned into the EcoR I and Not I sites of pcDNA 3.1 minigene Del-m ( , inset).

    Electrophoresis:

    Article Title: Cloning and molecular characterization of Δ12-fatty acid desaturase gene from Mortierella isabellina
    Article Snippet: .. PCR product about 1.2 kb long was gel-purified by electrophoresis and cloned into pGEM-T vector. .. Positive clones were selected on LB plate by color reaction.

    Polymerase Chain Reaction:

    Article Title: Engineering Saccharomyces cerevisiae for improvement in ethanol tolerance by accumulation of trehalose
    Article Snippet: .. The 5′ UP nth1 -kanMX-3′ DOWN nth1 , obtained from the third round of PCR, was ligated with pGEMT vector (Promega Corporation, Madison, WI, USA) and transformed into CaCl2 -treated E. coli TOP10F’ according to Hanahan and Meselson. .. The transformed host was screened by blue/white selection.

    Article Title: Octamer binding protein 2 (Oct2) regulates PD-L2 gene expression in B-1 cells through lineage-specific activity of a unique, intronic promoter
    Article Snippet: .. After checking the band pattern, PCR products were TA-cloned into pGEM-T vector (Promega, Madison, WI, USA). ..

    Article Title: Cloning and molecular characterization of Δ12-fatty acid desaturase gene from Mortierella isabellina
    Article Snippet: .. PCR product about 1.2 kb long was gel-purified by electrophoresis and cloned into pGEM-T vector. .. Positive clones were selected on LB plate by color reaction.

    Article Title: Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale
    Article Snippet: .. In the case of mixed infections (presence of double peaks in the analyzed pherograms from the sequencing of the original PCR amplicon), purified PCR products were ligated into pGEM-T easy cloning vector following the manufacturer's instructions (pGEM-T clone kit; Promega, Madison, WI) and transformed into Escherichia coli DH5α competent cells by heat shock. .. Positive colonies were selected by blue/white screening followed by colony PCR using M13F/R primers under the following conditions: 5 min at 95°C and 35 cycles of 60 s at 95°C, 60 s at 51°C, and 1 min and 20 s at 72°C and a final extension of 5 min at 72°C.

    Touchdown PCR:

    Article Title: Motifs within the CA-repeat-rich region of Surfactant Protein B (SFTPB) intron 4 differentially affect mRNA splicing
    Article Snippet: .. To improve the specificity of amplification in order to generate different length variants, the following touchdown PCR program ( ) was used: 95°C for 2 min, 5 cycles of 95°C for 30 s, 70°C for 30 s and 68°C for 30 s, then 5 cycles of 95°C for 30 s, 68°C for 30 s and 68°C for 30 s, and then 5 cycles of 95°C for 30 s, 66°C for 30 s and 68°C for 30 s, followed by 25 cycles of 94°C for 30 s, 64°C for 30 s and 68°C for 30 s and a final extension step at 68°C for 5 min. All PCR fragments, ranging from 96 to 437 were separated on agarose gel, purified and cloned into a pGEM-T vector (Promega, WI, USA). .. The recombinant plasmid DNAs were digested, purified and sub-cloned into the EcoR I and Not I sites of pcDNA 3.1 minigene Del-m ( , inset).

    Sequencing:

    Article Title: Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale
    Article Snippet: .. In the case of mixed infections (presence of double peaks in the analyzed pherograms from the sequencing of the original PCR amplicon), purified PCR products were ligated into pGEM-T easy cloning vector following the manufacturer's instructions (pGEM-T clone kit; Promega, Madison, WI) and transformed into Escherichia coli DH5α competent cells by heat shock. .. Positive colonies were selected by blue/white screening followed by colony PCR using M13F/R primers under the following conditions: 5 min at 95°C and 35 cycles of 60 s at 95°C, 60 s at 51°C, and 1 min and 20 s at 72°C and a final extension of 5 min at 72°C.

    Transformation Assay:

    Article Title: Engineering Saccharomyces cerevisiae for improvement in ethanol tolerance by accumulation of trehalose
    Article Snippet: .. The 5′ UP nth1 -kanMX-3′ DOWN nth1 , obtained from the third round of PCR, was ligated with pGEMT vector (Promega Corporation, Madison, WI, USA) and transformed into CaCl2 -treated E. coli TOP10F’ according to Hanahan and Meselson. .. The transformed host was screened by blue/white selection.

    Article Title: Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale
    Article Snippet: .. In the case of mixed infections (presence of double peaks in the analyzed pherograms from the sequencing of the original PCR amplicon), purified PCR products were ligated into pGEM-T easy cloning vector following the manufacturer's instructions (pGEM-T clone kit; Promega, Madison, WI) and transformed into Escherichia coli DH5α competent cells by heat shock. .. Positive colonies were selected by blue/white screening followed by colony PCR using M13F/R primers under the following conditions: 5 min at 95°C and 35 cycles of 60 s at 95°C, 60 s at 51°C, and 1 min and 20 s at 72°C and a final extension of 5 min at 72°C.

    Plasmid Preparation:

    Article Title: Chromosome-Based Genetic Complementation System for Xylella fastidiosa ▿
    Article Snippet: .. The resulting 1.6-kb fragment was then cloned into the pGEM-T vector (Promega), creating plasmid pAX1. .. PCR was also used to generate DNA fragments carrying the multiple cloning sites and the four antibiotic resistance cassettes that were inserted into pAX1.

    Article Title: Engineering Saccharomyces cerevisiae for improvement in ethanol tolerance by accumulation of trehalose
    Article Snippet: .. The 5′ UP nth1 -kanMX-3′ DOWN nth1 , obtained from the third round of PCR, was ligated with pGEMT vector (Promega Corporation, Madison, WI, USA) and transformed into CaCl2 -treated E. coli TOP10F’ according to Hanahan and Meselson. .. The transformed host was screened by blue/white selection.

    Article Title: Octamer binding protein 2 (Oct2) regulates PD-L2 gene expression in B-1 cells through lineage-specific activity of a unique, intronic promoter
    Article Snippet: .. After checking the band pattern, PCR products were TA-cloned into pGEM-T vector (Promega, Madison, WI, USA). ..

    Article Title: Cloning and molecular characterization of Δ12-fatty acid desaturase gene from Mortierella isabellina
    Article Snippet: .. PCR product about 1.2 kb long was gel-purified by electrophoresis and cloned into pGEM-T vector. .. Positive clones were selected on LB plate by color reaction.

    Article Title: Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale
    Article Snippet: .. In the case of mixed infections (presence of double peaks in the analyzed pherograms from the sequencing of the original PCR amplicon), purified PCR products were ligated into pGEM-T easy cloning vector following the manufacturer's instructions (pGEM-T clone kit; Promega, Madison, WI) and transformed into Escherichia coli DH5α competent cells by heat shock. .. Positive colonies were selected by blue/white screening followed by colony PCR using M13F/R primers under the following conditions: 5 min at 95°C and 35 cycles of 60 s at 95°C, 60 s at 51°C, and 1 min and 20 s at 72°C and a final extension of 5 min at 72°C.

    Article Title: Two RNA editing sites with cis-acting elements of moderate sequence identity are recognized by an identical site-recognition protein in tobacco chloroplasts
    Article Snippet: .. The amplified fragments were cloned into a pGEM-T vector using the pGEM-T Vector System (Promega). .. From these cloned plasmids, RNA substrates for in vitro editing and UV-crosslinking were prepared as previously described ( ) with slight modifications.

    Article Title: A gene encoding a protein modified by the phytohormone indoleacetic acid
    Article Snippet: .. The iap1 cDNA was cloned into a pGEM-T vector and was used for coupled in vitro transcription and translation using the TnT Quick Coupled System (Promega) with [35 S]methionine. .. GeneRacer Kit (Invitrogen) was used for full-length, RNA ligase-mediated rapid amplification of 5′ ends using RNA isolated 24 days after flowering.

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  • 93
    Promega pgem t easy cloning vector
    (A) Southern blotting of EcoRI-digested total DNA from FD-C-infected and FD-D-infected and healthy (H) periwinkles probed with a DIG-labeled malG gene amplicon obtained through <t>PCR</t> driven by primer pair malG _F/ malG _R (C+, probe positive control represented by <t>pGEM-T-</t> malG1 plasmid). (B) Electrophoresis separation of amplicons obtained following PCR of total DNA from FD-C-infected and FD-D-infected periwinkles with copy-specific primer pairs (002 and 005), according to the draft genome of FD92, and from healthy periwinkle. *, nonspecific PCR product.
    Pgem T Easy Cloning Vector, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 282 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy cloning vector/product/Promega
    Average 93 stars, based on 282 article reviews
    Price from $9.99 to $1999.99
    pgem t easy cloning vector - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    99
    Promega pgem t vector
    Identification of the recombinant plasmid, pTMICL12 by restriction enzyme. 1: <t>pGEM-T/EcoRI/XhoI;</t> 2: DNA size marker λ DNA/BstII; 3: pTMICL12/EcoRI/XhoI; 4: PCR product of D12D DNA.
    Pgem T Vector, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 2577 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t vector/product/Promega
    Average 99 stars, based on 2577 article reviews
    Price from $9.99 to $1999.99
    pgem t vector - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    (A) Southern blotting of EcoRI-digested total DNA from FD-C-infected and FD-D-infected and healthy (H) periwinkles probed with a DIG-labeled malG gene amplicon obtained through PCR driven by primer pair malG _F/ malG _R (C+, probe positive control represented by pGEM-T- malG1 plasmid). (B) Electrophoresis separation of amplicons obtained following PCR of total DNA from FD-C-infected and FD-D-infected periwinkles with copy-specific primer pairs (002 and 005), according to the draft genome of FD92, and from healthy periwinkle. *, nonspecific PCR product.

    Journal: Applied and Environmental Microbiology

    Article Title: Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale

    doi: 10.1128/AEM.03123-18

    Figure Lengend Snippet: (A) Southern blotting of EcoRI-digested total DNA from FD-C-infected and FD-D-infected and healthy (H) periwinkles probed with a DIG-labeled malG gene amplicon obtained through PCR driven by primer pair malG _F/ malG _R (C+, probe positive control represented by pGEM-T- malG1 plasmid). (B) Electrophoresis separation of amplicons obtained following PCR of total DNA from FD-C-infected and FD-D-infected periwinkles with copy-specific primer pairs (002 and 005), according to the draft genome of FD92, and from healthy periwinkle. *, nonspecific PCR product.

    Article Snippet: In the case of mixed infections (presence of double peaks in the analyzed pherograms from the sequencing of the original PCR amplicon), purified PCR products were ligated into pGEM-T easy cloning vector following the manufacturer's instructions (pGEM-T clone kit; Promega, Madison, WI) and transformed into Escherichia coli DH5α competent cells by heat shock.

    Techniques: Southern Blot, Infection, Labeling, Amplification, Polymerase Chain Reaction, Positive Control, Plasmid Preparation, Electrophoresis

    Identification of the recombinant plasmid, pTMICL12 by restriction enzyme. 1: pGEM-T/EcoRI/XhoI; 2: DNA size marker λ DNA/BstII; 3: pTMICL12/EcoRI/XhoI; 4: PCR product of D12D DNA.

    Journal:

    Article Title: Cloning and molecular characterization of Δ12-fatty acid desaturase gene from Mortierella isabellina

    doi: 10.3748/wjg.v12.i21.3373

    Figure Lengend Snippet: Identification of the recombinant plasmid, pTMICL12 by restriction enzyme. 1: pGEM-T/EcoRI/XhoI; 2: DNA size marker λ DNA/BstII; 3: pTMICL12/EcoRI/XhoI; 4: PCR product of D12D DNA.

    Article Snippet: PCR product about 1.2 kb long was gel-purified by electrophoresis and cloned into pGEM-T vector.

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

    WT intron 4 pGEM-T construct

    Journal: Journal of molecular biochemistry

    Article Title: Motifs within the CA-repeat-rich region of Surfactant Protein B (SFTPB) intron 4 differentially affect mRNA splicing

    doi:

    Figure Lengend Snippet: WT intron 4 pGEM-T construct

    Article Snippet: To improve the specificity of amplification in order to generate different length variants, the following touchdown PCR program ( ) was used: 95°C for 2 min, 5 cycles of 95°C for 30 s, 70°C for 30 s and 68°C for 30 s, then 5 cycles of 95°C for 30 s, 68°C for 30 s and 68°C for 30 s, and then 5 cycles of 95°C for 30 s, 66°C for 30 s and 68°C for 30 s, followed by 25 cycles of 94°C for 30 s, 64°C for 30 s and 68°C for 30 s and a final extension step at 68°C for 5 min. All PCR fragments, ranging from 96 to 437 were separated on agarose gel, purified and cloned into a pGEM-T vector (Promega, WI, USA).

    Techniques: Construct

    Disruption of BBA74 by insertion of kanamycin resistance cassette. (A) Schematic diagram of mutant construct. The region of lp54 between BBA73 and BBA76 was amplified by PCR using primers p73F/p76R and cloned into pGEM-T. The insertion of the kanamycin

    Journal:

    Article Title: Comparative Transcriptional Profiling of Borrelia burgdorferi Clinical Isolates Differing in Capacities for Hematogenous Dissemination

    doi: 10.1128/IAI.73.10.6791-6802.2005

    Figure Lengend Snippet: Disruption of BBA74 by insertion of kanamycin resistance cassette. (A) Schematic diagram of mutant construct. The region of lp54 between BBA73 and BBA76 was amplified by PCR using primers p73F/p76R and cloned into pGEM-T. The insertion of the kanamycin

    Article Snippet: A 2,327-bp DNA fragment spanning BBA73 to BBA76 (nucleotides 50601 to 52908 on plasmid lp54; GenBank accession number, ) was amplified by PCR using primers pA73F and pA76R (Table ) as forward and reverse primers, respectively, and cloned into the plasmid vector pGEM-T (Promega).

    Techniques: Mutagenesis, Construct, Amplification, Polymerase Chain Reaction, Clone Assay