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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: 88/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pgem t easy cloning vector/product/Promega
Average 88 stars, based on 16 article reviews
Price from $9.99 to $1999.99
pgem t easy cloning vector - by Bioz Stars, 2020-05
88/100 stars

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1) Product Images from "Genetic Diversity of Flavescence Dorée Phytoplasmas at the Vineyard Scale"

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

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.03123-18

(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.
Figure Legend 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.

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

Related Articles

Clone Assay:

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: Brain-Derived Neurotrophic Factor Is Critically Involved in Thermal-Experience-Dependent Developmental Plasticity
Article Snippet: .. The PCR product of 919 bp was subcloned into the pGEM-T Easy Cloning Vector (Promega, Madison, WI). .. Forward (F) and reverse (R) primers used to identify BDNF, NT-3, and NGF by reverse transcription (RT)-PCR were as follows: NGF, F, 5′-GGACCAAGAGGACTGCACAT-3′; NGF, R, 5′-GGGGGATAGGAGGATGTTGT-3′; NT-3, F, 5′-ACAGACCTCCTAAGGCAGCA-3′; NT-3, R, 5′-GGATGTCTTGCACTGGGAGT-3′; BDNF, F 5′-ACTGGCGGACACTTTTGAAC-3′; and BDNF, R, 5′-GTTGCACCAGACATGT-CCAC-3′.

Article Title: Brain-Derived Neurotrophic Factor Is Critically Involved in Thermal-Experience-Dependent Developmental Plasticity
Article Snippet: .. The PCR product was inserted into pGEM-T Easy cloning vector and was used as a template for probe synthesis. .. The hybridized sections were embedded in emulsion and developed after 30 d. The sections were stained with methyl green counterstain.

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.

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.

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.

Polymerase Chain 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: Brain-Derived Neurotrophic Factor Is Critically Involved in Thermal-Experience-Dependent Developmental Plasticity
Article Snippet: .. The PCR product of 919 bp was subcloned into the pGEM-T Easy Cloning Vector (Promega, Madison, WI). .. Forward (F) and reverse (R) primers used to identify BDNF, NT-3, and NGF by reverse transcription (RT)-PCR were as follows: NGF, F, 5′-GGACCAAGAGGACTGCACAT-3′; NGF, R, 5′-GGGGGATAGGAGGATGTTGT-3′; NT-3, F, 5′-ACAGACCTCCTAAGGCAGCA-3′; NT-3, R, 5′-GGATGTCTTGCACTGGGAGT-3′; BDNF, F 5′-ACTGGCGGACACTTTTGAAC-3′; and BDNF, R, 5′-GTTGCACCAGACATGT-CCAC-3′.

Article Title: Brain-Derived Neurotrophic Factor Is Critically Involved in Thermal-Experience-Dependent Developmental Plasticity
Article Snippet: .. The PCR product was inserted into pGEM-T Easy cloning vector and was used as a template for probe synthesis. .. The hybridized sections were embedded in emulsion and developed after 30 d. The sections were stained with methyl green counterstain.

Transformation Assay:

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: 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: Brain-Derived Neurotrophic Factor Is Critically Involved in Thermal-Experience-Dependent Developmental Plasticity
Article Snippet: .. The PCR product of 919 bp was subcloned into the pGEM-T Easy Cloning Vector (Promega, Madison, WI). .. Forward (F) and reverse (R) primers used to identify BDNF, NT-3, and NGF by reverse transcription (RT)-PCR were as follows: NGF, F, 5′-GGACCAAGAGGACTGCACAT-3′; NGF, R, 5′-GGGGGATAGGAGGATGTTGT-3′; NT-3, F, 5′-ACAGACCTCCTAAGGCAGCA-3′; NT-3, R, 5′-GGATGTCTTGCACTGGGAGT-3′; BDNF, F 5′-ACTGGCGGACACTTTTGAAC-3′; and BDNF, R, 5′-GTTGCACCAGACATGT-CCAC-3′.

Article Title: Brain-Derived Neurotrophic Factor Is Critically Involved in Thermal-Experience-Dependent Developmental Plasticity
Article Snippet: .. The PCR product was inserted into pGEM-T Easy cloning vector and was used as a template for probe synthesis. .. The hybridized sections were embedded in emulsion and developed after 30 d. The sections were stained with methyl green counterstain.

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    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: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy cloning vector/product/Promega
    Average 99 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    pgem t easy cloning vector - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

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    (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

    Amplification of the housekeeping genes and confirmation of cloning by EcoRI digestion of pGEM-T vector. (A) Amplification of the six target genes from cDNA of eggplant Lanes - M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (454 bp), 3: GAPDH (586bp), 4: Cyclophilin (265 bp), 5: Actin (333 bp), 6: RuBP (269bp). (B) Confirmation of the inserts in the recombinant pGEM-T vector by EcoRI digestion. Lanes - M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (454 bp), 3: Cyclophilin (265 bp), 4: GAPDH (586bp), 5: Actin (333 bp), 6: RuBP (269bp). (C) Amplification of the six target genes from genomic DNA of eggplant. Lanes- M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (No amplification), 3: Cyclophilin (265 bp), 4: GAPDH (1.2 kb), 5: Actin (333 bp), 6: RuBP (269bp).

    Journal: BMC Research Notes

    Article Title: Selection and validation of reference genes for quantitative gene expression studies by real-time PCR in eggplant (Solanum melongena L)

    doi: 10.1186/1756-0500-6-312

    Figure Lengend Snippet: Amplification of the housekeeping genes and confirmation of cloning by EcoRI digestion of pGEM-T vector. (A) Amplification of the six target genes from cDNA of eggplant Lanes - M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (454 bp), 3: GAPDH (586bp), 4: Cyclophilin (265 bp), 5: Actin (333 bp), 6: RuBP (269bp). (B) Confirmation of the inserts in the recombinant pGEM-T vector by EcoRI digestion. Lanes - M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (454 bp), 3: Cyclophilin (265 bp), 4: GAPDH (586bp), 5: Actin (333 bp), 6: RuBP (269bp). (C) Amplification of the six target genes from genomic DNA of eggplant. Lanes- M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (No amplification), 3: Cyclophilin (265 bp), 4: GAPDH (1.2 kb), 5: Actin (333 bp), 6: RuBP (269bp).

    Article Snippet: The amplified cDNA PCR products were cloned into pGEM-T Easy cloning vector (Promega) according to the manufacturer’s instructions.

    Techniques: Amplification, Clone Assay, Plasmid Preparation, Marker, Recombinant

    Rates of degradation of four different substrates by E. coli strains . Measurements were performed at 37°C, using one-dimensional 1 H-NMR spectroscopy (n = 3). Control: wild-type; pGUO0303: mutant transformed with pGEM-T EasyΩ cj0641 ; pGUO0304: mutant transformed with pGEM-T EasyΩ cj0774c ; pGUO0305: mutant transformed with pGEM-T EasyΩ cj1663 ; pGU0306: mutant transformed with pGEM-T EasyΩ cj064 1Ω Km R ; pGU0307: mutant transformed with pGEM-T EasyΩ cj0774c Ω Km R ; and pGU0308: mutant transformed with pGEM-T EasyΩ cj1663 Ω Km R . The activities of the wild-type were considered 100% and mutant activities are presented relative to the total control activity for the specific substrate. Histogram patterns correspond to degradation rates of PhePhn : inclined lines; PhnAce : horizontal lines; AmePhn : dots; and PhePhi : empty. The errors were calculated by determining the standard deviation from the mean of triplicate experiments.

    Journal: Gut Pathogens

    Article Title: Characterisation of Campylobacter jejuni genes potentially involved in phosphonate degradation

    doi: 10.1186/1757-4749-1-13

    Figure Lengend Snippet: Rates of degradation of four different substrates by E. coli strains . Measurements were performed at 37°C, using one-dimensional 1 H-NMR spectroscopy (n = 3). Control: wild-type; pGUO0303: mutant transformed with pGEM-T EasyΩ cj0641 ; pGUO0304: mutant transformed with pGEM-T EasyΩ cj0774c ; pGUO0305: mutant transformed with pGEM-T EasyΩ cj1663 ; pGU0306: mutant transformed with pGEM-T EasyΩ cj064 1Ω Km R ; pGU0307: mutant transformed with pGEM-T EasyΩ cj0774c Ω Km R ; and pGU0308: mutant transformed with pGEM-T EasyΩ cj1663 Ω Km R . The activities of the wild-type were considered 100% and mutant activities are presented relative to the total control activity for the specific substrate. Histogram patterns correspond to degradation rates of PhePhn : inclined lines; PhnAce : horizontal lines; AmePhn : dots; and PhePhi : empty. The errors were calculated by determining the standard deviation from the mean of triplicate experiments.

    Article Snippet: Cloning and screening of recombinant plasmids pGU0303, pGU0304 and pGU0305 Amplified PCR fragments were cloned into the bacterial cloning vector system pGEM-T Easy (Promega; Madison, WI, USA) in E. coli by standard cloning techniques [ ].

    Techniques: Nuclear Magnetic Resonance, Spectroscopy, Mutagenesis, Transformation Assay, Activity Assay, Standard Deviation

    PCR validation of alternative splicing events identified by Iso-Seq. cDNA from control (C) and treated (T) samples was used for PCR. Primer sets (F, forward and R, reverse) were designed to flank the splicing events. PCR products were excised from the gel, purified, cloned into pGEM-T Easy Vectors and sequenced from both directions. Sequences were aligned to the corresponding gene sequence and the structures of the novel isoforms were verified. Exons are represented by filled boxes, introns by lines and 3′ and 5′ UTRs are represented by open boxes. The gene models have one annotated isoform, which is shown in black. The novel isoforms that are supported by PacBio reads and/or sequencing of PCR products are colour-coded and indicated by arrows. Different splicing events are represented by lines connecting exons. Predicted protein for each isoform and putative domains predicted using the simple modular architecture tool (SMART) are presented in the right panel. The location of the predicted stop codon in the transcripts is represented by a vertical line. Alt. 3′, alternative acceptor site; Alt. 5′, alternative donor site; ES, exon skipping; IR, intron retention; SP, signal peptide; TM, transmembrane; M, lane with DNA size markers. Gene ID is shown at the left for each panel.

    Journal: Nature Communications

    Article Title: A survey of the sorghum transcriptome using single-molecule long reads

    doi: 10.1038/ncomms11706

    Figure Lengend Snippet: PCR validation of alternative splicing events identified by Iso-Seq. cDNA from control (C) and treated (T) samples was used for PCR. Primer sets (F, forward and R, reverse) were designed to flank the splicing events. PCR products were excised from the gel, purified, cloned into pGEM-T Easy Vectors and sequenced from both directions. Sequences were aligned to the corresponding gene sequence and the structures of the novel isoforms were verified. Exons are represented by filled boxes, introns by lines and 3′ and 5′ UTRs are represented by open boxes. The gene models have one annotated isoform, which is shown in black. The novel isoforms that are supported by PacBio reads and/or sequencing of PCR products are colour-coded and indicated by arrows. Different splicing events are represented by lines connecting exons. Predicted protein for each isoform and putative domains predicted using the simple modular architecture tool (SMART) are presented in the right panel. The location of the predicted stop codon in the transcripts is represented by a vertical line. Alt. 3′, alternative acceptor site; Alt. 5′, alternative donor site; ES, exon skipping; IR, intron retention; SP, signal peptide; TM, transmembrane; M, lane with DNA size markers. Gene ID is shown at the left for each panel.

    Article Snippet: The purified products were cloned onto pGEM-T Easy cloning vector (Promega) and sequenced from both directions using T7 and SP6 primers.

    Techniques: Polymerase Chain Reaction, Purification, Clone Assay, Sequencing