pgem-t easy vector Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Promega pgemt easy vector
    Localization of <t>Stx2</t> in brains of mice inoculated with pStx2. The area observed in this study is located in the mouse CA2 hippocampus. (A) Brain from control mouse inoculated with <t>pGEMT.</t> (B) Brain from mouse inoculated with 3 µg of pStx2 24 h after inoculation. (C) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation. (D) Brain from mouse injected with 0.5 ng of Stx2 4 days after injection. (E) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation, without primary antibody. (F) Number of Stx2-immunopositive cells in hippocampus per micrograph. Scale bar = 50 µm.
    Pgemt Easy Vector, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 15845 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgemt easy vector/product/Promega
    Average 99 stars, based on 15845 article reviews
    Price from $9.99 to $1999.99
    pgemt easy vector - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    85
    Promega pgem t easy vector cloning system
    Localization of <t>Stx2</t> in brains of mice inoculated with pStx2. The area observed in this study is located in the mouse CA2 hippocampus. (A) Brain from control mouse inoculated with <t>pGEMT.</t> (B) Brain from mouse inoculated with 3 µg of pStx2 24 h after inoculation. (C) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation. (D) Brain from mouse injected with 0.5 ng of Stx2 4 days after injection. (E) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation, without primary antibody. (F) Number of Stx2-immunopositive cells in hippocampus per micrograph. Scale bar = 50 µm.
    Pgem T Easy Vector Cloning System, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector cloning system/product/Promega
    Average 85 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector cloning system - by Bioz Stars, 2020-04
    85/100 stars
      Buy from Supplier

    92
    Millipore pgem t easy vector
    Localization of <t>Stx2</t> in brains of mice inoculated with pStx2. The area observed in this study is located in the mouse CA2 hippocampus. (A) Brain from control mouse inoculated with <t>pGEMT.</t> (B) Brain from mouse inoculated with 3 µg of pStx2 24 h after inoculation. (C) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation. (D) Brain from mouse injected with 0.5 ng of Stx2 4 days after injection. (E) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation, without primary antibody. (F) Number of Stx2-immunopositive cells in hippocampus per micrograph. Scale bar = 50 µm.
    Pgem T Easy Vector, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 192 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/Millipore
    Average 92 stars, based on 192 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-04
    92/100 stars
      Buy from Supplier

    94
    Thermo Fisher pgem t easy vector
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 970 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/Thermo Fisher
    Average 94 stars, based on 970 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    97
    TaKaRa pgem t easy vector
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 97/100, based on 343 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/TaKaRa
    Average 97 stars, based on 343 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    87
    Bio-Rad pgem t easy vector
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 87/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/Bio-Rad
    Average 87 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    87
    Stratagene pgem t easy vector
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector, supplied by Stratagene, used in various techniques. Bioz Stars score: 87/100, based on 134 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/Stratagene
    Average 87 stars, based on 134 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    88
    Sangon Biotech pgem t easy vector
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector, supplied by Sangon Biotech, used in various techniques. Bioz Stars score: 88/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/Sangon Biotech
    Average 88 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-04
    88/100 stars
      Buy from Supplier

    94
    tiangen biotech co pgem t easy vector
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 94/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/tiangen biotech co
    Average 94 stars, based on 62 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    87
    TaKaRa pgem t easy vector system
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector System, supplied by TaKaRa, used in various techniques. Bioz Stars score: 87/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector system/product/TaKaRa
    Average 87 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector system - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    93
    Thermo Fisher pgem t easy vector system
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector system/product/Thermo Fisher
    Average 93 stars, based on 133 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector system - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    Image Search Results


    Localization of Stx2 in brains of mice inoculated with pStx2. The area observed in this study is located in the mouse CA2 hippocampus. (A) Brain from control mouse inoculated with pGEMT. (B) Brain from mouse inoculated with 3 µg of pStx2 24 h after inoculation. (C) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation. (D) Brain from mouse injected with 0.5 ng of Stx2 4 days after injection. (E) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation, without primary antibody. (F) Number of Stx2-immunopositive cells in hippocampus per micrograph. Scale bar = 50 µm.

    Journal: mBio

    Article Title: Promoter Sequence of Shiga Toxin 2 (Stx2) Is Recognized In Vivo, Leading to Production of Biologically Active Stx2

    doi: 10.1128/mBio.00501-13

    Figure Lengend Snippet: Localization of Stx2 in brains of mice inoculated with pStx2. The area observed in this study is located in the mouse CA2 hippocampus. (A) Brain from control mouse inoculated with pGEMT. (B) Brain from mouse inoculated with 3 µg of pStx2 24 h after inoculation. (C) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation. (D) Brain from mouse injected with 0.5 ng of Stx2 4 days after injection. (E) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation, without primary antibody. (F) Number of Stx2-immunopositive cells in hippocampus per micrograph. Scale bar = 50 µm.

    Article Snippet: The pGEMT Easy vector carrying the sequence of Stx2 was digested and religated in order to delete the active site of Stx2, generating the plasmid pStx2ΔAB ( ) as a Stx2-specific biological activity control.

    Techniques: Mouse Assay, Injection

    (A) Mice displayed a lethal dose-related response to pStx2. Survival rates of mice injected with different doses of pStx2 are shown. Mice injected with pStx2ΔAB or pGEMT were used as negative controls. Mice injected with 1 LD 100 and 3 LD 100 of purified Stx2 were used as positive controls. (B) Stx2 mRNA was detected in the liver. A real-time-PCR curve using cDNA from livers of mice inoculated with pStx2 (dotted line) or pGEMT (gray line) is shown. As a positive control, 50 pg of plasmid pStx2 (bold line) was used. (C) DNA electrophoresis. Lane 1, 50-bp ladder; lane 2. cDNA from livers of mice inoculated with pStx2; lane 3, cDNA from livers of mice inoculated with pGEMT; lane 4, pStx2 was used as positive control.

    Journal: mBio

    Article Title: Promoter Sequence of Shiga Toxin 2 (Stx2) Is Recognized In Vivo, Leading to Production of Biologically Active Stx2

    doi: 10.1128/mBio.00501-13

    Figure Lengend Snippet: (A) Mice displayed a lethal dose-related response to pStx2. Survival rates of mice injected with different doses of pStx2 are shown. Mice injected with pStx2ΔAB or pGEMT were used as negative controls. Mice injected with 1 LD 100 and 3 LD 100 of purified Stx2 were used as positive controls. (B) Stx2 mRNA was detected in the liver. A real-time-PCR curve using cDNA from livers of mice inoculated with pStx2 (dotted line) or pGEMT (gray line) is shown. As a positive control, 50 pg of plasmid pStx2 (bold line) was used. (C) DNA electrophoresis. Lane 1, 50-bp ladder; lane 2. cDNA from livers of mice inoculated with pStx2; lane 3, cDNA from livers of mice inoculated with pGEMT; lane 4, pStx2 was used as positive control.

    Article Snippet: The pGEMT Easy vector carrying the sequence of Stx2 was digested and religated in order to delete the active site of Stx2, generating the plasmid pStx2ΔAB ( ) as a Stx2-specific biological activity control.

    Techniques: Mouse Assay, Injection, Purification, Real-time Polymerase Chain Reaction, Positive Control, Plasmid Preparation, Nucleic Acid Electrophoresis

    Construction of transgenic C. elegans expressing Venus under the control of T21D12.3 enhancer/promoter. T21D12.3 and T21D12.4 ( pat-6 ) are transcriptionally regulated as an operon. Therefore, 6.7 kb genome DNA from the transcriptional initiation site to the neighboring gene to pat-6 was subcloned into pGEMT-T plasmid. By injecting the plasmid into germ line of adult worm, we obtained transgenic nematodes in which the Venus cDNA was subcloned into exon 4 to express a PQBP1-Venus fusion protein under the control of endogenous enhancer/promoter.

    Journal: PLoS ONE

    Article Title: Nematode Homologue of PQBP1, a Mental Retardation Causative Gene, Is Involved in Lipid Metabolism

    doi: 10.1371/journal.pone.0004104

    Figure Lengend Snippet: Construction of transgenic C. elegans expressing Venus under the control of T21D12.3 enhancer/promoter. T21D12.3 and T21D12.4 ( pat-6 ) are transcriptionally regulated as an operon. Therefore, 6.7 kb genome DNA from the transcriptional initiation site to the neighboring gene to pat-6 was subcloned into pGEMT-T plasmid. By injecting the plasmid into germ line of adult worm, we obtained transgenic nematodes in which the Venus cDNA was subcloned into exon 4 to express a PQBP1-Venus fusion protein under the control of endogenous enhancer/promoter.

    Article Snippet: The genome fragment was subcloned into pGEMT-T Easy vector (Promega) and the resultant plasmid (pGEMT-T-T21D12.3) was digested with Pml I (BioLabs) which is unique in the exon 4 of the T21D12.3 gene.

    Techniques: Transgenic Assay, Expressing, Plasmid Preparation

    Electrophoresis result of recombinant plasmid pGEM-T Easy-Mpt83 (P1-P6).

    Journal: Journal of Genetic Engineering & Biotechnology

    Article Title: Cloning and expression of MPT83 gene from Mycobacterium tuberculosis in E. coli BL21 as vaccine candidate of tuberculosis: A preliminary study

    doi: 10.1016/j.jgeb.2018.04.001

    Figure Lengend Snippet: Electrophoresis result of recombinant plasmid pGEM-T Easy-Mpt83 (P1-P6).

    Article Snippet: The cloning of MPT83 gene into pGEM-T Easy vector results in recombinant plasmid with the size of 3678 bp.

    Techniques: Electrophoresis, Recombinant, Plasmid Preparation

    E. coli growth pattern in LB-Amp plus IPTG and X-Gal Plate containing pGEM-T Easy-Mpt83 recombinant plasmid.

    Journal: Journal of Genetic Engineering & Biotechnology

    Article Title: Cloning and expression of MPT83 gene from Mycobacterium tuberculosis in E. coli BL21 as vaccine candidate of tuberculosis: A preliminary study

    doi: 10.1016/j.jgeb.2018.04.001

    Figure Lengend Snippet: E. coli growth pattern in LB-Amp plus IPTG and X-Gal Plate containing pGEM-T Easy-Mpt83 recombinant plasmid.

    Article Snippet: The cloning of MPT83 gene into pGEM-T Easy vector results in recombinant plasmid with the size of 3678 bp.

    Techniques: Recombinant, Plasmid Preparation

    pELMO and pGEM-T Easy vector cloning efficiency for different sized inserts. pELMO and pGEM-T Easy cloning efficiency regarding low, medium and large sized inserts. Cloning efficiency is expressed as the ratio of the amount of PCR positive colonies

    Journal: AMB Express

    Article Title: pELMO, an optimised in-house cloning vector

    doi: 10.1186/s13568-017-0324-2

    Figure Lengend Snippet: pELMO and pGEM-T Easy vector cloning efficiency for different sized inserts. pELMO and pGEM-T Easy cloning efficiency regarding low, medium and large sized inserts. Cloning efficiency is expressed as the ratio of the amount of PCR positive colonies

    Article Snippet: Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Techniques: Plasmid Preparation, Clone Assay, Polymerase Chain Reaction

    Agarose gel electrophoresis (1.2%) of the pGEM-T/CTRL-1 cDNA and pGEM-T/CTRL-1 genomic DNA after EcoR I digestion. M: 100-bp size marker; lane 1: full-length CTRL-1 cDNA PCR product; lane 2: pGEM-T/CTRL-1 cDNA; lane 3: CTRL-1 genomic DNA PCR product; lane 4: pGEM-T/CTRL-1 genomic DNA.

    Journal: Toxins

    Article Title: Cloning a Chymotrypsin-Like 1 (CTRL-1) Protease cDNA from the Jellyfish Nemopilema nomurai

    doi: 10.3390/toxins8070205

    Figure Lengend Snippet: Agarose gel electrophoresis (1.2%) of the pGEM-T/CTRL-1 cDNA and pGEM-T/CTRL-1 genomic DNA after EcoR I digestion. M: 100-bp size marker; lane 1: full-length CTRL-1 cDNA PCR product; lane 2: pGEM-T/CTRL-1 cDNA; lane 3: CTRL-1 genomic DNA PCR product; lane 4: pGEM-T/CTRL-1 genomic DNA.

    Article Snippet: All the PCR products were purified with the Expin™ GeneAll® PCR SV purification kit (GeneAll, Seoul, Korea), cloned into the pGEM®-T Easy Vector System (Promega, Madison, WI, USA) and confirmed the clone by EcoR I digestion at 37 °C for 1 h. Full-length of cDNA sequence was identified by ABI PRISM 3739 Genetic Analyzer (Thermo Fisher, Waltham, MA, USA).

    Techniques: Agarose Gel Electrophoresis, Marker, Polymerase Chain Reaction

    Schematic representation of the capsule genes of W135 serogroup in transcriptional fusion of synG with ermAM cassette . The synG gene responsible for the synthesis of the W135 capsule was amplified using the 98-30 and 03-12-5 oligonucleotides (Table 3) from W135 ATCC strain. The amplified fragment was cloned into the pGEM-T Easy Vector System I (Promega, Madison, WI, USA), to generate the plasmid pLAN11. In the same conditions, another fragment was amplified using the 04.02-2/galECK29A from synG downstream sequence to generate pLAN52. The ermAM cassette was insered into Nco I site of pLAN52 to generate pLAN53. The fragment amplified from pLAN53 with the ERAM1 and galECK29A (Dolan Livengood [ 23 ] et al ., 2003) was insered into Pst I site of pLAN11 to generate pLAN13-2. This plasmid was linearised by the enzyme SphI and transformed into W135 ATCC strain to generate the synG::ermAM strain M6, erythromycin resistant.

    Journal: Journal of Nanobiotechnology

    Article Title: Effect of mesoporous silica under Neisseria meningitidis transformation process: environmental effects under meningococci transformation

    doi: 10.1186/1477-3155-9-28

    Figure Lengend Snippet: Schematic representation of the capsule genes of W135 serogroup in transcriptional fusion of synG with ermAM cassette . The synG gene responsible for the synthesis of the W135 capsule was amplified using the 98-30 and 03-12-5 oligonucleotides (Table 3) from W135 ATCC strain. The amplified fragment was cloned into the pGEM-T Easy Vector System I (Promega, Madison, WI, USA), to generate the plasmid pLAN11. In the same conditions, another fragment was amplified using the 04.02-2/galECK29A from synG downstream sequence to generate pLAN52. The ermAM cassette was insered into Nco I site of pLAN52 to generate pLAN53. The fragment amplified from pLAN53 with the ERAM1 and galECK29A (Dolan Livengood [ 23 ] et al ., 2003) was insered into Pst I site of pLAN11 to generate pLAN13-2. This plasmid was linearised by the enzyme SphI and transformed into W135 ATCC strain to generate the synG::ermAM strain M6, erythromycin resistant.

    Article Snippet: The amplified fragment was cloned into the pGEM-T Easy Vector System I (Promega, Madison, WI, USA), to generate the plasmid pLAN11.

    Techniques: Amplification, Clone Assay, Plasmid Preparation, Sequencing, Transformation Assay

    Schematic representation of the capsule genes of C serogroup in disrupted construction of NMB0065 gene with aaDA cassette . The NMB0065 gene was amplified using the 03-12-3 and 03-12-4 oligonucleotides (Table 3) from C2135 strain. This fragment was cloned into the pGEM-T Easy Vector System II (Promega Corporation, Madison, WI, USA), to generate the plasmid pLAN6. E. coli strain Z501 was transformed with plasmid pLAN6 resulting in the plasmid pLAN7. The Ω aaDA cassette was inserted into the BclI site of pLAN7 to generate plasmid pLAN45, which was transformed into the C2135 strain to generate the isogenic mutant strain M2.

    Journal: Journal of Nanobiotechnology

    Article Title: Effect of mesoporous silica under Neisseria meningitidis transformation process: environmental effects under meningococci transformation

    doi: 10.1186/1477-3155-9-28

    Figure Lengend Snippet: Schematic representation of the capsule genes of C serogroup in disrupted construction of NMB0065 gene with aaDA cassette . The NMB0065 gene was amplified using the 03-12-3 and 03-12-4 oligonucleotides (Table 3) from C2135 strain. This fragment was cloned into the pGEM-T Easy Vector System II (Promega Corporation, Madison, WI, USA), to generate the plasmid pLAN6. E. coli strain Z501 was transformed with plasmid pLAN6 resulting in the plasmid pLAN7. The Ω aaDA cassette was inserted into the BclI site of pLAN7 to generate plasmid pLAN45, which was transformed into the C2135 strain to generate the isogenic mutant strain M2.

    Article Snippet: This fragment was cloned into the pGEM-T Easy Vector System II (Promega Corporation, Madison, WI, USA), to generate the plasmid pLAN6.

    Techniques: Amplification, Clone Assay, Plasmid Preparation, Transformation Assay, Mutagenesis

    Strategies for generation of dsRNAs and d-siRNAs. (A) PCR template strategy was utilized for obtaining s GFP , An rasA and An ras B dsRNA in a single T7 transcription reaction. T7 promoter sequence was added to both forward and reverse gene specific primers of s GFP , An rasA and An rasB , and then PCR amplification was performed in order to generate templates for dsRNA synthesis. (B) For generation of template DNA for unrelated MiAchE , PCR amplification was performed on pGEM T- MiAchE vector using M13 primers. This amplification includes the T7 promoter to one end and SP6 promoter to another end of MiAchE . (C) PCR templates for transcription of respective target genes. M. ladder; B. blank; s GFP , An rasA , An rasB and MiAchE target gene templates. (D) Synthesis of dsRNAs with T7 or SP6 in vitro transcription reactions. M. Ladder; s GFP , An rasA , An rasB and MiAchE dsRNAs. (E) 20% PAGE analysis of purified diced siRNAs. M-Ladder, s GFP , An rasA , An rasB and MiAchE d-siRNAs. The d-siRNAs of all target genes were generated by cleaving respective dsRNAs with RNase III at 37°C for 30 mins, followed by subsequent purifications and finally dissolved in nuclease free water.

    Journal: PLoS ONE

    Article Title: Targeting Fungal Genes by Diced siRNAs: A Rapid Tool to Decipher Gene Function in Aspergillus nidulans

    doi: 10.1371/journal.pone.0075443

    Figure Lengend Snippet: Strategies for generation of dsRNAs and d-siRNAs. (A) PCR template strategy was utilized for obtaining s GFP , An rasA and An ras B dsRNA in a single T7 transcription reaction. T7 promoter sequence was added to both forward and reverse gene specific primers of s GFP , An rasA and An rasB , and then PCR amplification was performed in order to generate templates for dsRNA synthesis. (B) For generation of template DNA for unrelated MiAchE , PCR amplification was performed on pGEM T- MiAchE vector using M13 primers. This amplification includes the T7 promoter to one end and SP6 promoter to another end of MiAchE . (C) PCR templates for transcription of respective target genes. M. ladder; B. blank; s GFP , An rasA , An rasB and MiAchE target gene templates. (D) Synthesis of dsRNAs with T7 or SP6 in vitro transcription reactions. M. Ladder; s GFP , An rasA , An rasB and MiAchE dsRNAs. (E) 20% PAGE analysis of purified diced siRNAs. M-Ladder, s GFP , An rasA , An rasB and MiAchE d-siRNAs. The d-siRNAs of all target genes were generated by cleaving respective dsRNAs with RNase III at 37°C for 30 mins, followed by subsequent purifications and finally dissolved in nuclease free water.

    Article Snippet: The AnrasA and AnrasB PCR products were purified using PCR clean up system (Fermentas, USA) and cloned in pGEM-T easy (Promega, USA) vector and confirmed by automated DNA sequencing.

    Techniques: Polymerase Chain Reaction, Sequencing, Amplification, Plasmid Preparation, In Vitro, Polyacrylamide Gel Electrophoresis, Purification, Generated

    Construction of full-length cDNA clone of VHSV DK-3952B genome. Six overlapping cDNA fragments covering the entire VHSV genome were assembled by ligation into a pCI vector using the EcoRI, PvuII, EcoRV, KpnI, SalI, BclI, SmaI, StuI, BsrGI and NotI restriction sites. Assembly was carried out in 3 steps; (i) cloning of six fragments (F1-F6) separately in pGEM-T vectors (see methods), and addition of hammerhead ribozyme (HHRz) at the 5′-end of F1 and hepatitis delta ribozyme (HdvRz) at the 3′-end of F6; (ii) construction of A, B and C clones into the pCI vector; (iii) assembly of the full-length clone by ligating fragments from these three clones in the pCI vector. Restriction sites artificially created (*) and naturally present in the genome are indicated. Abbreviation: CMV, cytomegalovirus immediate-early enhancer and promoter

    Journal: Virology Journal

    Article Title: The glycoprotein, non-virion protein, and polymerase of viral hemorrhagic septicemia virus are not determinants of host-specific virulence in rainbow trout

    doi: 10.1186/s12985-019-1139-3

    Figure Lengend Snippet: Construction of full-length cDNA clone of VHSV DK-3952B genome. Six overlapping cDNA fragments covering the entire VHSV genome were assembled by ligation into a pCI vector using the EcoRI, PvuII, EcoRV, KpnI, SalI, BclI, SmaI, StuI, BsrGI and NotI restriction sites. Assembly was carried out in 3 steps; (i) cloning of six fragments (F1-F6) separately in pGEM-T vectors (see methods), and addition of hammerhead ribozyme (HHRz) at the 5′-end of F1 and hepatitis delta ribozyme (HdvRz) at the 3′-end of F6; (ii) construction of A, B and C clones into the pCI vector; (iii) assembly of the full-length clone by ligating fragments from these three clones in the pCI vector. Restriction sites artificially created (*) and naturally present in the genome are indicated. Abbreviation: CMV, cytomegalovirus immediate-early enhancer and promoter

    Article Snippet: The cDNA fragments obtained after RT-PCR amplification were cloned into pGEM-T or pGEM-T Easy vectors (Promega Corp., Madison, WI).

    Techniques: Ligation, Plasmid Preparation, Clone Assay

    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the pGEM-T vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).

    Journal: Molecular Vision

    Article Title: FGFR2 molecular analysis and related clinical findings in one Chinese family with Crouzon Syndrome

    doi:

    Figure Lengend Snippet: DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the pGEM-T vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).

    Article Snippet: The purified PCR fragments were ligated into the pGEM-T easy vector (Invitrogen, Carlsbad, CA), and the resulting plasmids were transfected, by heat shock, into DH5a-competent Escherichia coli for propagation.

    Techniques: Sequencing, Mutagenesis, Plasmid Preparation