pgem t easy  (Promega)

 
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  • 99
    Name:
    pGEM T Easy Vector Systems
    Description:
    PCR cloning vectors with 3 options for insert excision
    Catalog Number:
    a1360
    Price:
    None
    Category:
    Nucleic Acid Extraction Analysis PCR PCR Cloning
    Buy from Supplier


    Structured Review

    Promega pgem t easy
    PCR cloning vectors with 3 options for insert excision
    https://www.bioz.com/result/pgem t easy/product/Promega
    Average 99 stars, based on 2417 article reviews
    Price from $9.99 to $1999.99
    pgem t easy - by Bioz Stars, 2020-01
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: The anthelmintic drug praziquantel activates a schistosome transient receptor potential channel
    Article Snippet: Molecular cloning For cloning of Sm .TRPMPZQ , total RNA was isolated from adult schistosome worm pairs using TRIzol® and poly(A)-purified using a NucleoTrap® mRNA minikit. cDNA was synthesized using the SuperScriptTM III first-strand synthesis system (Invitrogen). .. Using the predicted sequence (Smp_246790) as a template, cDNA from transcribed sequences was amplified by PCR (LA TaqTM polymerase) and ligated into pGEM®-T Easy (Promega) for sequencing.

    Article Title: Variable responses of formyl peptide receptor haplotypes toward bacterial peptides
    Article Snippet: Paragraph title: Cloning of FPR haplotypes ... Haplotypes were identified after ligation of the PCR amplicons into pGEM® -T Easy (Promega, Inc., Madison, WI), and sequencing of the plasmid-insert with T7 and SP6 primers.

    Article Title: Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation
    Article Snippet: For YFP fragment complementation assays, PRC-amplified Eed and Ezh2 fragments were cloned into split YFP expression vectors . .. Briefly, Eed PCR products were TA-cloned into pGEM®-T Easy (Promega) and then subcloned into the Not I and Cla I sites at the 5′ end of Venus 2.

    Article Title: Bacterial biota in reflux esophagitis and Barrett's esophagus
    Article Snippet: Paragraph title: Cloning and sequencing ... The PCR products were separated from free PCR primers using a PCR purification kit (Qiagen), then ligated with the pGEM® T Easy (Promega) vector, and used to transform into E coli DH5α competent cells.

    Article Title: Biosynthesis of heme O in intraerythrocytic stages of Plasmodium falciparum and potential inhibitors of this pathway
    Article Snippet: .. Vector construction To obtain the transfection vector containing the desired cox10 gene of P. falciparum , the cox10 ORF was PCR-amplified (primers 5′-agatctATGGGATTTAATAAGATTTTTCC and 5′-ctgcagcTTTAAATGTTCTTTTTATCAAATGTAGG, program 94 °C, 40 s, 54 °C, 40 s, 65 °C 1 min 30 s, 30 cycles) on genomic DNA from PF3d7, using Elongase enzyme mix (Thermo Scientific/Invitrogen, Carlsbad, CA, USA) and cloned into pGEM® T-easy (Promega, Wisconsin, USA). .. The fragment was sequenced and a fragment containing the correct sequence was excised using via the introduced Bgl II and Pst I sites and ligated in the pRESA-GFP/HA vector and cut with the same enzymes.

    Article Title: Capsular glucan and intracellular glycogen of Mycobacterium tuberculosis: Biosynthesis and Impact on the Persistence in mice
    Article Snippet: Sal I-cut resulted in the deletion of 740 bp of the coding sequence of glgB . glgB::Km was then cloned with the xylE colored marker ( ) into the BamH I site of pJQ200 to yield pJQ glgBKX , the construct used for allelic replacement at the glgB locus. .. The glgC gene and flanking regions was amplified using primers C364.1 (5’-cccgaattcggctgggatagacccgcaac-3’) and C364.2 (5’-cccgaattcggcgtttcatcagcagttcg-3’) and inserted into pGEM®-T easy (Promega), yielding pGEMT glgC .

    Functional Assay:

    Article Title: The anthelmintic drug praziquantel activates a schistosome transient receptor potential channel
    Article Snippet: Using the predicted sequence (Smp_246790) as a template, cDNA from transcribed sequences was amplified by PCR (LA TaqTM polymerase) and ligated into pGEM®-T Easy (Promega) for sequencing. .. The sequence used here for functional analyses represents the reference sequence (2268 amino acids, Smp_246790.5).

    Amplification:

    Article Title: The anthelmintic drug praziquantel activates a schistosome transient receptor potential channel
    Article Snippet: .. Using the predicted sequence (Smp_246790) as a template, cDNA from transcribed sequences was amplified by PCR (LA TaqTM polymerase) and ligated into pGEM®-T Easy (Promega) for sequencing. ..

    Article Title: Variable responses of formyl peptide receptor haplotypes toward bacterial peptides
    Article Snippet: Amplification utilized primer pairs FPR-41F22 and FPR+1063R22, where the first nucleotide of the primer is indicated by its position relative to the adenosine in the ATG translation initiation site, followed by F or R for forward or reverse and the number of nucleotides in the primer. .. Haplotypes were identified after ligation of the PCR amplicons into pGEM® -T Easy (Promega, Inc., Madison, WI), and sequencing of the plasmid-insert with T7 and SP6 primers.

    Article Title: Capsular glucan and intracellular glycogen of Mycobacterium tuberculosis: Biosynthesis and Impact on the Persistence in mice
    Article Snippet: .. The glgC gene and flanking regions was amplified using primers C364.1 (5’-cccgaattcggctgggatagacccgcaac-3’) and C364.2 (5’-cccgaattcggcgtttcatcagcagttcg-3’) and inserted into pGEM®-T easy (Promega), yielding pGEMT glgC . .. A disrupted allele of the glgC gene was then constructed by cloning the Km cassette into the BamH I site of glgC . glgC::Km was finally cloned with the xylE gene into the BamH I site of pPR27 , to yield p27 glgCKX . treZ was amplified using the couple of primers B48.2 (5’-gcttcctgggcggcgcataccatc-3’) and B48.3 (5’-cccgaattcggccggctccgcagcccgcag-3’) and a disrupted allele of the treZ gene was constructed by cloning the Km cassette into the Xho I site of treZ . treZ::Km was then cloned with the xylE gene into the BamH I site of pPR27, yielding p27 treZKX .

    Fluorescence:

    Article Title: Biosynthesis of heme O in intraerythrocytic stages of Plasmodium falciparum and potential inhibitors of this pathway
    Article Snippet: Vector construction To obtain the transfection vector containing the desired cox10 gene of P. falciparum , the cox10 ORF was PCR-amplified (primers 5′-agatctATGGGATTTAATAAGATTTTTCC and 5′-ctgcagcTTTAAATGTTCTTTTTATCAAATGTAGG, program 94 °C, 40 s, 54 °C, 40 s, 65 °C 1 min 30 s, 30 cycles) on genomic DNA from PF3d7, using Elongase enzyme mix (Thermo Scientific/Invitrogen, Carlsbad, CA, USA) and cloned into pGEM® T-easy (Promega, Wisconsin, USA). .. Integrated parasites were checked for green fluorescence.

    Molecular Cloning:

    Article Title: The anthelmintic drug praziquantel activates a schistosome transient receptor potential channel
    Article Snippet: Paragraph title: Molecular cloning ... Using the predicted sequence (Smp_246790) as a template, cDNA from transcribed sequences was amplified by PCR (LA TaqTM polymerase) and ligated into pGEM®-T Easy (Promega) for sequencing.

    Isolation:

    Article Title: The anthelmintic drug praziquantel activates a schistosome transient receptor potential channel
    Article Snippet: Molecular cloning For cloning of Sm .TRPMPZQ , total RNA was isolated from adult schistosome worm pairs using TRIzol® and poly(A)-purified using a NucleoTrap® mRNA minikit. cDNA was synthesized using the SuperScriptTM III first-strand synthesis system (Invitrogen). .. Using the predicted sequence (Smp_246790) as a template, cDNA from transcribed sequences was amplified by PCR (LA TaqTM polymerase) and ligated into pGEM®-T Easy (Promega) for sequencing.

    Article Title: Variable responses of formyl peptide receptor haplotypes toward bacterial peptides
    Article Snippet: Genomic DNA was isolated from healthy donors from 250 μl whole blood using E.Z.N.A. .. Haplotypes were identified after ligation of the PCR amplicons into pGEM® -T Easy (Promega, Inc., Madison, WI), and sequencing of the plasmid-insert with T7 and SP6 primers.

    Subcloning:

    Article Title: Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation
    Article Snippet: Briefly, Eed PCR products were TA-cloned into pGEM®-T Easy (Promega) and then subcloned into the Not I and Cla I sites at the 5′ end of Venus 2. .. Similarly, PCR-amplified Ezh2 fragments were TA-cloned before subcloning into the the Bsp EI and Xba I sites at the 3′ end of Venus 1.

    Synthesized:

    Article Title: The anthelmintic drug praziquantel activates a schistosome transient receptor potential channel
    Article Snippet: Molecular cloning For cloning of Sm .TRPMPZQ , total RNA was isolated from adult schistosome worm pairs using TRIzol® and poly(A)-purified using a NucleoTrap® mRNA minikit. cDNA was synthesized using the SuperScriptTM III first-strand synthesis system (Invitrogen). .. Using the predicted sequence (Smp_246790) as a template, cDNA from transcribed sequences was amplified by PCR (LA TaqTM polymerase) and ligated into pGEM®-T Easy (Promega) for sequencing.

    Ligation:

    Article Title: Variable responses of formyl peptide receptor haplotypes toward bacterial peptides
    Article Snippet: .. Haplotypes were identified after ligation of the PCR amplicons into pGEM® -T Easy (Promega, Inc., Madison, WI), and sequencing of the plasmid-insert with T7 and SP6 primers. ..

    Construct:

    Article Title: Biosynthesis of heme O in intraerythrocytic stages of Plasmodium falciparum and potential inhibitors of this pathway
    Article Snippet: Vector construction To obtain the transfection vector containing the desired cox10 gene of P. falciparum , the cox10 ORF was PCR-amplified (primers 5′-agatctATGGGATTTAATAAGATTTTTCC and 5′-ctgcagcTTTAAATGTTCTTTTTATCAAATGTAGG, program 94 °C, 40 s, 54 °C, 40 s, 65 °C 1 min 30 s, 30 cycles) on genomic DNA from PF3d7, using Elongase enzyme mix (Thermo Scientific/Invitrogen, Carlsbad, CA, USA) and cloned into pGEM® T-easy (Promega, Wisconsin, USA). .. The construct was transfected into P. falciparum 3D7 as described .

    Article Title: Capsular glucan and intracellular glycogen of Mycobacterium tuberculosis: Biosynthesis and Impact on the Persistence in mice
    Article Snippet: Sal I-cut resulted in the deletion of 740 bp of the coding sequence of glgB . glgB::Km was then cloned with the xylE colored marker ( ) into the BamH I site of pJQ200 to yield pJQ glgBKX , the construct used for allelic replacement at the glgB locus. .. The glgC gene and flanking regions was amplified using primers C364.1 (5’-cccgaattcggctgggatagacccgcaac-3’) and C364.2 (5’-cccgaattcggcgtttcatcagcagttcg-3’) and inserted into pGEM®-T easy (Promega), yielding pGEMT glgC .

    Purification:

    Article Title: Bacterial biota in reflux esophagitis and Barrett's esophagus
    Article Snippet: .. The PCR products were separated from free PCR primers using a PCR purification kit (Qiagen), then ligated with the pGEM® T Easy (Promega) vector, and used to transform into E coli DH5α competent cells. .. The cloned inserts underwent sequence analysis using vector-based primers.

    Sequencing:

    Article Title: The anthelmintic drug praziquantel activates a schistosome transient receptor potential channel
    Article Snippet: .. Using the predicted sequence (Smp_246790) as a template, cDNA from transcribed sequences was amplified by PCR (LA TaqTM polymerase) and ligated into pGEM®-T Easy (Promega) for sequencing. ..

    Article Title: Variable responses of formyl peptide receptor haplotypes toward bacterial peptides
    Article Snippet: .. Haplotypes were identified after ligation of the PCR amplicons into pGEM® -T Easy (Promega, Inc., Madison, WI), and sequencing of the plasmid-insert with T7 and SP6 primers. ..

    Article Title: Bacterial biota in reflux esophagitis and Barrett's esophagus
    Article Snippet: Paragraph title: Cloning and sequencing ... The PCR products were separated from free PCR primers using a PCR purification kit (Qiagen), then ligated with the pGEM® T Easy (Promega) vector, and used to transform into E coli DH5α competent cells.

    Article Title: Biosynthesis of heme O in intraerythrocytic stages of Plasmodium falciparum and potential inhibitors of this pathway
    Article Snippet: Vector construction To obtain the transfection vector containing the desired cox10 gene of P. falciparum , the cox10 ORF was PCR-amplified (primers 5′-agatctATGGGATTTAATAAGATTTTTCC and 5′-ctgcagcTTTAAATGTTCTTTTTATCAAATGTAGG, program 94 °C, 40 s, 54 °C, 40 s, 65 °C 1 min 30 s, 30 cycles) on genomic DNA from PF3d7, using Elongase enzyme mix (Thermo Scientific/Invitrogen, Carlsbad, CA, USA) and cloned into pGEM® T-easy (Promega, Wisconsin, USA). .. The fragment was sequenced and a fragment containing the correct sequence was excised using via the introduced Bgl II and Pst I sites and ligated in the pRESA-GFP/HA vector and cut with the same enzymes.

    Article Title: Capsular glucan and intracellular glycogen of Mycobacterium tuberculosis: Biosynthesis and Impact on the Persistence in mice
    Article Snippet: Sal I-cut resulted in the deletion of 740 bp of the coding sequence of glgB . glgB::Km was then cloned with the xylE colored marker ( ) into the BamH I site of pJQ200 to yield pJQ glgBKX , the construct used for allelic replacement at the glgB locus. .. The glgC gene and flanking regions was amplified using primers C364.1 (5’-cccgaattcggctgggatagacccgcaac-3’) and C364.2 (5’-cccgaattcggcgtttcatcagcagttcg-3’) and inserted into pGEM®-T easy (Promega), yielding pGEMT glgC .

    Polymerase Chain Reaction:

    Article Title: The anthelmintic drug praziquantel activates a schistosome transient receptor potential channel
    Article Snippet: .. Using the predicted sequence (Smp_246790) as a template, cDNA from transcribed sequences was amplified by PCR (LA TaqTM polymerase) and ligated into pGEM®-T Easy (Promega) for sequencing. ..

    Article Title: Variable responses of formyl peptide receptor haplotypes toward bacterial peptides
    Article Snippet: .. Haplotypes were identified after ligation of the PCR amplicons into pGEM® -T Easy (Promega, Inc., Madison, WI), and sequencing of the plasmid-insert with T7 and SP6 primers. ..

    Article Title: Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation
    Article Snippet: .. Briefly, Eed PCR products were TA-cloned into pGEM®-T Easy (Promega) and then subcloned into the Not I and Cla I sites at the 5′ end of Venus 2. .. Similarly, PCR-amplified Ezh2 fragments were TA-cloned before subcloning into the the Bsp EI and Xba I sites at the 3′ end of Venus 1.

    Article Title: Bacterial biota in reflux esophagitis and Barrett's esophagus
    Article Snippet: .. The PCR products were separated from free PCR primers using a PCR purification kit (Qiagen), then ligated with the pGEM® T Easy (Promega) vector, and used to transform into E coli DH5α competent cells. .. The cloned inserts underwent sequence analysis using vector-based primers.

    Article Title: Biosynthesis of heme O in intraerythrocytic stages of Plasmodium falciparum and potential inhibitors of this pathway
    Article Snippet: .. Vector construction To obtain the transfection vector containing the desired cox10 gene of P. falciparum , the cox10 ORF was PCR-amplified (primers 5′-agatctATGGGATTTAATAAGATTTTTCC and 5′-ctgcagcTTTAAATGTTCTTTTTATCAAATGTAGG, program 94 °C, 40 s, 54 °C, 40 s, 65 °C 1 min 30 s, 30 cycles) on genomic DNA from PF3d7, using Elongase enzyme mix (Thermo Scientific/Invitrogen, Carlsbad, CA, USA) and cloned into pGEM® T-easy (Promega, Wisconsin, USA). .. The fragment was sequenced and a fragment containing the correct sequence was excised using via the introduced Bgl II and Pst I sites and ligated in the pRESA-GFP/HA vector and cut with the same enzymes.

    Article Title: Capsular glucan and intracellular glycogen of Mycobacterium tuberculosis: Biosynthesis and Impact on the Persistence in mice
    Article Snippet: Standard PCR strategies were used to amplify the M. tuberculosis H37Rv treZ, glgB, glgA, Rv3032 and glgC genes. glgA and its flanking regions were amplified using primers glgA.1 (5’-gcggaattccgcggtcgcattttcacgtgg-3’) and glgA.2 (5’-gcggaattcggatggtcgacgaccatatcc-3’), and a disrupted allele of the glgA gene was constructed by cloning the kanamycin (Km) resistance cassette from pUC4K (Amersham Pharmacia Biotech) into the Nco I sites of glgA . .. The glgC gene and flanking regions was amplified using primers C364.1 (5’-cccgaattcggctgggatagacccgcaac-3’) and C364.2 (5’-cccgaattcggcgtttcatcagcagttcg-3’) and inserted into pGEM®-T easy (Promega), yielding pGEMT glgC .

    Generated:

    Article Title: Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation
    Article Snippet: Site-directed point mutations were generated by standard methods. .. Briefly, Eed PCR products were TA-cloned into pGEM®-T Easy (Promega) and then subcloned into the Not I and Cla I sites at the 5′ end of Venus 2.

    Transfection:

    Article Title: Biosynthesis of heme O in intraerythrocytic stages of Plasmodium falciparum and potential inhibitors of this pathway
    Article Snippet: .. Vector construction To obtain the transfection vector containing the desired cox10 gene of P. falciparum , the cox10 ORF was PCR-amplified (primers 5′-agatctATGGGATTTAATAAGATTTTTCC and 5′-ctgcagcTTTAAATGTTCTTTTTATCAAATGTAGG, program 94 °C, 40 s, 54 °C, 40 s, 65 °C 1 min 30 s, 30 cycles) on genomic DNA from PF3d7, using Elongase enzyme mix (Thermo Scientific/Invitrogen, Carlsbad, CA, USA) and cloned into pGEM® T-easy (Promega, Wisconsin, USA). .. The fragment was sequenced and a fragment containing the correct sequence was excised using via the introduced Bgl II and Pst I sites and ligated in the pRESA-GFP/HA vector and cut with the same enzymes.

    Expressing:

    Article Title: Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation
    Article Snippet: For YFP fragment complementation assays, PRC-amplified Eed and Ezh2 fragments were cloned into split YFP expression vectors . .. Briefly, Eed PCR products were TA-cloned into pGEM®-T Easy (Promega) and then subcloned into the Not I and Cla I sites at the 5′ end of Venus 2.

    Marker:

    Article Title: Capsular glucan and intracellular glycogen of Mycobacterium tuberculosis: Biosynthesis and Impact on the Persistence in mice
    Article Snippet: Sal I-cut resulted in the deletion of 740 bp of the coding sequence of glgB . glgB::Km was then cloned with the xylE colored marker ( ) into the BamH I site of pJQ200 to yield pJQ glgBKX , the construct used for allelic replacement at the glgB locus. .. The glgC gene and flanking regions was amplified using primers C364.1 (5’-cccgaattcggctgggatagacccgcaac-3’) and C364.2 (5’-cccgaattcggcgtttcatcagcagttcg-3’) and inserted into pGEM®-T easy (Promega), yielding pGEMT glgC .

    Plasmid Preparation:

    Article Title: Variable responses of formyl peptide receptor haplotypes toward bacterial peptides
    Article Snippet: .. Haplotypes were identified after ligation of the PCR amplicons into pGEM® -T Easy (Promega, Inc., Madison, WI), and sequencing of the plasmid-insert with T7 and SP6 primers. ..

    Article Title: Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation
    Article Snippet: Paragraph title: Plasmid Construction ... Briefly, Eed PCR products were TA-cloned into pGEM®-T Easy (Promega) and then subcloned into the Not I and Cla I sites at the 5′ end of Venus 2.

    Article Title: Bacterial biota in reflux esophagitis and Barrett's esophagus
    Article Snippet: .. The PCR products were separated from free PCR primers using a PCR purification kit (Qiagen), then ligated with the pGEM® T Easy (Promega) vector, and used to transform into E coli DH5α competent cells. .. The cloned inserts underwent sequence analysis using vector-based primers.

    Article Title: Biosynthesis of heme O in intraerythrocytic stages of Plasmodium falciparum and potential inhibitors of this pathway
    Article Snippet: .. Vector construction To obtain the transfection vector containing the desired cox10 gene of P. falciparum , the cox10 ORF was PCR-amplified (primers 5′-agatctATGGGATTTAATAAGATTTTTCC and 5′-ctgcagcTTTAAATGTTCTTTTTATCAAATGTAGG, program 94 °C, 40 s, 54 °C, 40 s, 65 °C 1 min 30 s, 30 cycles) on genomic DNA from PF3d7, using Elongase enzyme mix (Thermo Scientific/Invitrogen, Carlsbad, CA, USA) and cloned into pGEM® T-easy (Promega, Wisconsin, USA). .. The fragment was sequenced and a fragment containing the correct sequence was excised using via the introduced Bgl II and Pst I sites and ligated in the pRESA-GFP/HA vector and cut with the same enzymes.

    Article Title: Capsular glucan and intracellular glycogen of Mycobacterium tuberculosis: Biosynthesis and Impact on the Persistence in mice
    Article Snippet: Digestion by Nco I resulted in the deletion of 253 bp of the coding sequence of glgA . glgA::Km was then cloned into pJQ200- xylE , yielding pJQ glgAKX , the plasmid used for allelic replacement. .. The glgC gene and flanking regions was amplified using primers C364.1 (5’-cccgaattcggctgggatagacccgcaac-3’) and C364.2 (5’-cccgaattcggcgtttcatcagcagttcg-3’) and inserted into pGEM®-T easy (Promega), yielding pGEMT glgC .

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  • Bioz Stars
  • Bioz vStars
  • 99
    Promega pgem t easy
    Pgem T Easy, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 2417 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy/product/Promega
    Average 99 stars, based on 2417 article reviews
    Price from $9.99 to $1999.99
    pgem t easy - by Bioz Stars, 2020-01
    99/100 stars
      Buy from Supplier

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