ncoi  (Promega)

 
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    Name:
    NcoI
    Description:
    Restriction enzyme that cuts DNA at 37°C and leaves a 5 overhang
    Catalog Number:
    r6513
    Price:
    None
    Category:
    Nucleic Acid Extraction Analysis Cloning DNA Markers Restriction Enzymes
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    Structured Review

    Promega ncoi
    Detection of wild-type, <t>Asp299Gly,</t> and Thr399Ile alleles in the human TLR4 gene. Genomic DNA of individuals recruited into the current study was amplified according to the method of Lorenz et al. (reference 4 ). PCR products were digested with the enzymes <t>NcoI</t> (299 allele) or HinfI (399 allele). Lanes 1 and 14, 100 base pair marker. Lanes 2–7, RFLP of the 299 section of TLR4 gene from three wild-type donors and three heterozygotes. Lanes 8–13, RFLP of the 399 section of TLR4 gene from three wild-type donors and three heterozygotes. Top band represents presence of wild-type allele, bottom band indicates presence of mutant allele. Genotypes were confirmed by sequencing.
    Restriction enzyme that cuts DNA at 37°C and leaves a 5 overhang
    https://www.bioz.com/result/ncoi/product/Promega
    Average 94 stars, based on 118 article reviews
    Price from $9.99 to $1999.99
    ncoi - by Bioz Stars, 2020-07
    94/100 stars

    Images

    1) Product Images from "Monocytes Heterozygous for the Asp299Gly and Thr399Ile Mutations in the Toll-like Receptor 4 Gene Show No Deficit in Lipopolysaccharide Signalling"

    Article Title: Monocytes Heterozygous for the Asp299Gly and Thr399Ile Mutations in the Toll-like Receptor 4 Gene Show No Deficit in Lipopolysaccharide Signalling

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20022078

    Detection of wild-type, Asp299Gly, and Thr399Ile alleles in the human TLR4 gene. Genomic DNA of individuals recruited into the current study was amplified according to the method of Lorenz et al. (reference 4 ). PCR products were digested with the enzymes NcoI (299 allele) or HinfI (399 allele). Lanes 1 and 14, 100 base pair marker. Lanes 2–7, RFLP of the 299 section of TLR4 gene from three wild-type donors and three heterozygotes. Lanes 8–13, RFLP of the 399 section of TLR4 gene from three wild-type donors and three heterozygotes. Top band represents presence of wild-type allele, bottom band indicates presence of mutant allele. Genotypes were confirmed by sequencing.
    Figure Legend Snippet: Detection of wild-type, Asp299Gly, and Thr399Ile alleles in the human TLR4 gene. Genomic DNA of individuals recruited into the current study was amplified according to the method of Lorenz et al. (reference 4 ). PCR products were digested with the enzymes NcoI (299 allele) or HinfI (399 allele). Lanes 1 and 14, 100 base pair marker. Lanes 2–7, RFLP of the 299 section of TLR4 gene from three wild-type donors and three heterozygotes. Lanes 8–13, RFLP of the 399 section of TLR4 gene from three wild-type donors and three heterozygotes. Top band represents presence of wild-type allele, bottom band indicates presence of mutant allele. Genotypes were confirmed by sequencing.

    Techniques Used: Amplification, Polymerase Chain Reaction, Marker, Mutagenesis, Sequencing

    Related Articles

    Clone Assay:

    Article Title: RC1339/APRc from Rickettsia conorii Is a Novel Aspartic Protease with Properties of Retropepsin-Like Enzymes
    Article Snippet: .. The gene encoding the full-length APRc (construct coding amino acids 1–231) was then amplified to include restriction sites for NcoI and NotI at 5′- and 3′-ends, respectively, using the forward primer 5′-CCATGGGAATGAACAAAAAACTGATCAAACTG-3′ and the reverse primer 5′-CTCGAGATAATTCAGAATCAGCAGATCTTT-3′ ; the resulting PCR product was cloned into pGEM-T Easy plasmid (Promega). .. After digestion with NcoI and NotI, APRc1–231 insert was subcloned into pET28a expression vector (Invitrogen) in frame with a C-terminal His-tag (pET-APRc1–231 His).

    Amplification:

    Article Title: Identification and Characterization of CCAAT Enhancer-binding Protein (C/EBP) as a Transcriptional Activator for Epstein-Barr Virus Oncogene Latent Membrane Protein 1 *
    Article Snippet: .. The amplified DNA was digested with XhoI and NcoI, and then inserted into the XhoI/NcoI sites of pGL4.10 (Promega). .. Likewise, luciferase reporter constructs containing various TR sequences were prepared using the following primers: for pLMP1/ED-L1+TR-L1-FLuc, ED-L1p+TR-L1pFor, and ED-L1p+TR-L1pRev, for pLMP1/TR-L1-FLuc, TR-L1pFor, and TR-L1pRev, for pLMP1/TR-L1+BS-FLuc, TR-L1p+BSFor, and TR-L1p+BSRev ( ).

    Article Title: RC1339/APRc from Rickettsia conorii Is a Novel Aspartic Protease with Properties of Retropepsin-Like Enzymes
    Article Snippet: .. The gene encoding the full-length APRc (construct coding amino acids 1–231) was then amplified to include restriction sites for NcoI and NotI at 5′- and 3′-ends, respectively, using the forward primer 5′-CCATGGGAATGAACAAAAAACTGATCAAACTG-3′ and the reverse primer 5′-CTCGAGATAATTCAGAATCAGCAGATCTTT-3′ ; the resulting PCR product was cloned into pGEM-T Easy plasmid (Promega). .. After digestion with NcoI and NotI, APRc1–231 insert was subcloned into pET28a expression vector (Invitrogen) in frame with a C-terminal His-tag (pET-APRc1–231 His).

    Construct:

    Article Title: RC1339/APRc from Rickettsia conorii Is a Novel Aspartic Protease with Properties of Retropepsin-Like Enzymes
    Article Snippet: .. The gene encoding the full-length APRc (construct coding amino acids 1–231) was then amplified to include restriction sites for NcoI and NotI at 5′- and 3′-ends, respectively, using the forward primer 5′-CCATGGGAATGAACAAAAAACTGATCAAACTG-3′ and the reverse primer 5′-CTCGAGATAATTCAGAATCAGCAGATCTTT-3′ ; the resulting PCR product was cloned into pGEM-T Easy plasmid (Promega). .. After digestion with NcoI and NotI, APRc1–231 insert was subcloned into pET28a expression vector (Invitrogen) in frame with a C-terminal His-tag (pET-APRc1–231 His).

    Article Title: Human SREBP1c Expression in Liver Is Directly Regulated by Peroxisome Proliferator-activated Receptor ? (PPAR?) *
    Article Snippet: .. A 1564-bp fragment was obtained by NcoI digestion and subcloned in the NcoI site of pGL3-basic luciferase vector (Promega) to construct the −1564/+1-luc vector. .. The −520/+1-luc vector was prepared by PCR from the −1564/+1-luc vector using the forward primer 5′-GGAGGGTACCAGGCTCGCTCAGGGTGCCAGC-3′ and the reverse primer GLprimer2 (Promega) to be then inserted into the KpnI/NcoI site of the pGL3-basic vector.

    Luciferase:

    Article Title: Human SREBP1c Expression in Liver Is Directly Regulated by Peroxisome Proliferator-activated Receptor ? (PPAR?) *
    Article Snippet: .. A 1564-bp fragment was obtained by NcoI digestion and subcloned in the NcoI site of pGL3-basic luciferase vector (Promega) to construct the −1564/+1-luc vector. .. The −520/+1-luc vector was prepared by PCR from the −1564/+1-luc vector using the forward primer 5′-GGAGGGTACCAGGCTCGCTCAGGGTGCCAGC-3′ and the reverse primer GLprimer2 (Promega) to be then inserted into the KpnI/NcoI site of the pGL3-basic vector.

    Polymerase Chain Reaction:

    Article Title: Two polymorphisms of the tumour necrosis factor gene do not influence survival in pancreatic cancer
    Article Snippet: .. The PCR product was digested directly with 1 U of NcoI restriction enzyme (Promega, Madison, WI) at 37°C for 4 h. Restriction enzyme products were analysed on 1% NuSieve agarose (FMC Bioproducts, Rockland, ME) or 6% polyacrylamide gels (BioRad, Hemel Hempstead, UK). .. The cleaved product produced bands at 133 and 235 bp representing the allele TNFB1, while the uncleaved 368-bp product represented the allele TNFB2 ( ).

    Article Title: Two polymorphisms of the tumour necrosis factor gene do not influence survival in pancreatic cancer
    Article Snippet: .. The PCR product was digested directly with 1 U of NcoI restriction enzyme (Promega) at 37°C for 4 h. Restriction enzyme products were analysed on 9% polyacrylamide gels (BioRad). .. The cleaved product produced bands at 87 and 20 bp representing the TNF-308 A allele, while the uncleaved 107-bp product represented the TNF-308 G allele ( ).

    Article Title: RC1339/APRc from Rickettsia conorii Is a Novel Aspartic Protease with Properties of Retropepsin-Like Enzymes
    Article Snippet: .. The gene encoding the full-length APRc (construct coding amino acids 1–231) was then amplified to include restriction sites for NcoI and NotI at 5′- and 3′-ends, respectively, using the forward primer 5′-CCATGGGAATGAACAAAAAACTGATCAAACTG-3′ and the reverse primer 5′-CTCGAGATAATTCAGAATCAGCAGATCTTT-3′ ; the resulting PCR product was cloned into pGEM-T Easy plasmid (Promega). .. After digestion with NcoI and NotI, APRc1–231 insert was subcloned into pET28a expression vector (Invitrogen) in frame with a C-terminal His-tag (pET-APRc1–231 His).

    Article Title: Monocytes Heterozygous for the Asp299Gly and Thr399Ile Mutations in the Toll-like Receptor 4 Gene Show No Deficit in Lipopolysaccharide Signalling
    Article Snippet: .. Thermal cycling was: 95°C for 4 min, then 30 cycles of 95°C for 30 s, 55°C for 30 s, and 72°C for 30 s. Asp299Gly or Thr399Ile PCR products (20 μl) were digested with NcoI (Promega) or HinfI (Promega), respectively, and visualized on a 3% high resolution agar gel (Nusieve 3.1; Flowgen). ..

    Plasmid Preparation:

    Article Title: The ASK1 and ASK2 Genes Are Essential for Arabidopsis Early Development
    Article Snippet: .. Vectors were digested with NotI and transcribed with T7 RNA polymerase for both ASK1 and ASK2 antisense probes, whereas for the ASK1 sense probe, vector was digested with NcoI and transcribed with SP6 RNA polymerase (Promega). .. Histochemical staining for GUS activity was performed using the standard X-Gluc solution (100 mM sodium phosphate buffer, pH 7.0, 0.1% Triton X-100, 1 mg/mL 5 bromo-4-chloro-3-indolyl β- d -glucuronide [X-Gluc]) with the addition of either 2 or 10 mM potassium ferricyanide and potassium ferrocyanide.

    Article Title: RC1339/APRc from Rickettsia conorii Is a Novel Aspartic Protease with Properties of Retropepsin-Like Enzymes
    Article Snippet: .. The gene encoding the full-length APRc (construct coding amino acids 1–231) was then amplified to include restriction sites for NcoI and NotI at 5′- and 3′-ends, respectively, using the forward primer 5′-CCATGGGAATGAACAAAAAACTGATCAAACTG-3′ and the reverse primer 5′-CTCGAGATAATTCAGAATCAGCAGATCTTT-3′ ; the resulting PCR product was cloned into pGEM-T Easy plasmid (Promega). .. After digestion with NcoI and NotI, APRc1–231 insert was subcloned into pET28a expression vector (Invitrogen) in frame with a C-terminal His-tag (pET-APRc1–231 His).

    Article Title: Human SREBP1c Expression in Liver Is Directly Regulated by Peroxisome Proliferator-activated Receptor ? (PPAR?) *
    Article Snippet: .. A 1564-bp fragment was obtained by NcoI digestion and subcloned in the NcoI site of pGL3-basic luciferase vector (Promega) to construct the −1564/+1-luc vector. .. The −520/+1-luc vector was prepared by PCR from the −1564/+1-luc vector using the forward primer 5′-GGAGGGTACCAGGCTCGCTCAGGGTGCCAGC-3′ and the reverse primer GLprimer2 (Promega) to be then inserted into the KpnI/NcoI site of the pGL3-basic vector.

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  • 94
    Promega ncoi
    Detection of wild-type, <t>Asp299Gly,</t> and Thr399Ile alleles in the human TLR4 gene. Genomic DNA of individuals recruited into the current study was amplified according to the method of Lorenz et al. (reference 4 ). PCR products were digested with the enzymes <t>NcoI</t> (299 allele) or HinfI (399 allele). Lanes 1 and 14, 100 base pair marker. Lanes 2–7, RFLP of the 299 section of TLR4 gene from three wild-type donors and three heterozygotes. Lanes 8–13, RFLP of the 399 section of TLR4 gene from three wild-type donors and three heterozygotes. Top band represents presence of wild-type allele, bottom band indicates presence of mutant allele. Genotypes were confirmed by sequencing.
    Ncoi, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ncoi/product/Promega
    Average 94 stars, based on 124 article reviews
    Price from $9.99 to $1999.99
    ncoi - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    90
    Promega ncoi site
    Translational activation mediated by the 55mer 5′-UTR structure. ( A ) Plasmids carrying the 55mer (with bases 9–10 flipped to avoid a cryptic translational start site as these positions were essentially not conserved at all across the 14 genes, Figure 5A bases 9–10) at two different positions (the <t>HindIII</t> positions the 55mer close to the transcription start site whereas the <t>NcoI</t> positions the 55mer close to the translational start site with a distance of ∼30 nt from the two sites) were transfected into NIH 3T3 and NIH 3T3/4E cells, which were serum starved or maintained in normal media. The cells were harvested after 16 h and relative protein levels were measured using the dual luciferase assay. Shown are the ratios between serum starved and serum replete cells. Each experiment was normalized to the basic vector pGL3 control and the experiment was repeated three times in duplicate (two different constructs showed similar results and were pooled). (Error bars indicate standard deviation, *students t -test P -value
    Ncoi Site, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ncoi site/product/Promega
    Average 90 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    ncoi site - by Bioz Stars, 2020-07
    90/100 stars
      Buy from Supplier

    Image Search Results


    Detection of wild-type, Asp299Gly, and Thr399Ile alleles in the human TLR4 gene. Genomic DNA of individuals recruited into the current study was amplified according to the method of Lorenz et al. (reference 4 ). PCR products were digested with the enzymes NcoI (299 allele) or HinfI (399 allele). Lanes 1 and 14, 100 base pair marker. Lanes 2–7, RFLP of the 299 section of TLR4 gene from three wild-type donors and three heterozygotes. Lanes 8–13, RFLP of the 399 section of TLR4 gene from three wild-type donors and three heterozygotes. Top band represents presence of wild-type allele, bottom band indicates presence of mutant allele. Genotypes were confirmed by sequencing.

    Journal: The Journal of Experimental Medicine

    Article Title: Monocytes Heterozygous for the Asp299Gly and Thr399Ile Mutations in the Toll-like Receptor 4 Gene Show No Deficit in Lipopolysaccharide Signalling

    doi: 10.1084/jem.20022078

    Figure Lengend Snippet: Detection of wild-type, Asp299Gly, and Thr399Ile alleles in the human TLR4 gene. Genomic DNA of individuals recruited into the current study was amplified according to the method of Lorenz et al. (reference 4 ). PCR products were digested with the enzymes NcoI (299 allele) or HinfI (399 allele). Lanes 1 and 14, 100 base pair marker. Lanes 2–7, RFLP of the 299 section of TLR4 gene from three wild-type donors and three heterozygotes. Lanes 8–13, RFLP of the 399 section of TLR4 gene from three wild-type donors and three heterozygotes. Top band represents presence of wild-type allele, bottom band indicates presence of mutant allele. Genotypes were confirmed by sequencing.

    Article Snippet: Thermal cycling was: 95°C for 4 min, then 30 cycles of 95°C for 30 s, 55°C for 30 s, and 72°C for 30 s. Asp299Gly or Thr399Ile PCR products (20 μl) were digested with NcoI (Promega) or HinfI (Promega), respectively, and visualized on a 3% high resolution agar gel (Nusieve 3.1; Flowgen).

    Techniques: Amplification, Polymerase Chain Reaction, Marker, Mutagenesis, Sequencing

    TNFB Nco1 polymorphism. The polymerase chain reaction (PCR) was used to amplify a 368-bp fragment of the of the TNFβ genomic sequence and the PCR product was digested directly with 1 U of NcoI restriction enzyme. Lane 1 shows a homozygous for the cleaved product produced bands at 133 and 235 bp (with incomplete digestion of the 368-bp band) representing the allele TNFB1, while the uncleaved 368-bp product in lane 3 represents the allele TNFB2. Lane 2 shows a heterozygote pattern.

    Journal: Clinical and Experimental Immunology

    Article Title: Two polymorphisms of the tumour necrosis factor gene do not influence survival in pancreatic cancer

    doi: 10.1046/j.1365-2249.1999.01005.x

    Figure Lengend Snippet: TNFB Nco1 polymorphism. The polymerase chain reaction (PCR) was used to amplify a 368-bp fragment of the of the TNFβ genomic sequence and the PCR product was digested directly with 1 U of NcoI restriction enzyme. Lane 1 shows a homozygous for the cleaved product produced bands at 133 and 235 bp (with incomplete digestion of the 368-bp band) representing the allele TNFB1, while the uncleaved 368-bp product in lane 3 represents the allele TNFB2. Lane 2 shows a heterozygote pattern.

    Article Snippet: The PCR product was digested directly with 1 U of NcoI restriction enzyme (Promega, Madison, WI) at 37°C for 4 h. Restriction enzyme products were analysed on 1% NuSieve agarose (FMC Bioproducts, Rockland, ME) or 6% polyacrylamide gels (BioRad, Hemel Hempstead, UK).

    Techniques: Polymerase Chain Reaction, Sequencing, Produced

    Translational activation mediated by the 55mer 5′-UTR structure. ( A ) Plasmids carrying the 55mer (with bases 9–10 flipped to avoid a cryptic translational start site as these positions were essentially not conserved at all across the 14 genes, Figure 5A bases 9–10) at two different positions (the HindIII positions the 55mer close to the transcription start site whereas the NcoI positions the 55mer close to the translational start site with a distance of ∼30 nt from the two sites) were transfected into NIH 3T3 and NIH 3T3/4E cells, which were serum starved or maintained in normal media. The cells were harvested after 16 h and relative protein levels were measured using the dual luciferase assay. Shown are the ratios between serum starved and serum replete cells. Each experiment was normalized to the basic vector pGL3 control and the experiment was repeated three times in duplicate (two different constructs showed similar results and were pooled). (Error bars indicate standard deviation, *students t -test P -value

    Journal: Nucleic Acids Research

    Article Title: Apoptosis resistance downstream of eIF4E: posttranscriptional activation of an anti-apoptotic transcript carrying a consensus hairpin structure

    doi: 10.1093/nar/gkl558

    Figure Lengend Snippet: Translational activation mediated by the 55mer 5′-UTR structure. ( A ) Plasmids carrying the 55mer (with bases 9–10 flipped to avoid a cryptic translational start site as these positions were essentially not conserved at all across the 14 genes, Figure 5A bases 9–10) at two different positions (the HindIII positions the 55mer close to the transcription start site whereas the NcoI positions the 55mer close to the translational start site with a distance of ∼30 nt from the two sites) were transfected into NIH 3T3 and NIH 3T3/4E cells, which were serum starved or maintained in normal media. The cells were harvested after 16 h and relative protein levels were measured using the dual luciferase assay. Shown are the ratios between serum starved and serum replete cells. Each experiment was normalized to the basic vector pGL3 control and the experiment was repeated three times in duplicate (two different constructs showed similar results and were pooled). (Error bars indicate standard deviation, *students t -test P -value

    Article Snippet: Functional studies of novel mRNA elements The 5′ stem–loop was cloned into both the HindIII (close to the transcription start site) and the NcoI site (close to the translation start site) of the pGL3 control vector from Promega.

    Techniques: Activation Assay, Transfection, Luciferase, Plasmid Preparation, Construct, Standard Deviation