pcii  (New England Biolabs)


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    Name:
    PciI
    Description:
    PciI 1 000 units
    Catalog Number:
    r0655l
    Price:
    290
    Size:
    1 000 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs pcii
    PciI
    PciI 1 000 units
    https://www.bioz.com/result/pcii/product/New England Biolabs
    Average 94 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    pcii - by Bioz Stars, 2020-09
    94/100 stars

    Images

    1) Product Images from "Functional impacts of 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxycytosine at a single hemi-modified CpG dinucleotide in a gene promoter"

    Article Title: Functional impacts of 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxycytosine at a single hemi-modified CpG dinucleotide in a gene promoter

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkx718

    Impact of CpG hemi-methylation in the minimal CRE promoter on the CREB binding and the reporter EGFP gene expression. ( A ) Artificial CRE-uno promoter: transcription start (broken arrow), CRE sequence (bold), AatII site (underlined), Nb.BsrDI nicking positions (vertical arrows), and positions of 5-mC/5-hmC/5-fC/5-caC in the incorporated synthetic oligonucleotides (asterisks). ( B ) Inhibition of the AatII restriction endonuclease by the M.SssI methylation of the central CG-dinucleotide in CRE. Agarose gel electrophoresis of pCRE-uno incubated with AatII. Arrows indicate migration positions of the linearized vector (4408 bp) and of the covalently closed (cc) and nicked (nc) forms of circular vector DNA. ( C ) Verification of the incorporation of synthetic oligonucleotides containing one or three 5-mC (1 × 5mC, 3 × 5mC) by inhibition of the AatII cleavage. ( D ) Scheme of the PciI/BmtI digest of the pCRE-uno vector and electrophoretic mobility shift assay (EMSA) of the PciI/BmtI fragments under the indicated CREB:DNA molar ratios. The shorter (180 bp) fragment contains CRE with no or one 5-mC in the central CG-dinucleotide. The right panel shows control CRE-less vector (pCRE-zero) incubated with CREB in parallel. ( E ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one (in the central CG-dinucleotide of CRE) or three 5mC. Representative FACS data and relative EGFP expression values (mean ± SD, n = 7). Expression in the absence of an EGFP-coding vector (DsRed) denotes the lower detection limit. CRE-less vector (pCRE-zero) was included in two experiments as a reference for the basal expression level. P -values calculated by the Student's t -test.
    Figure Legend Snippet: Impact of CpG hemi-methylation in the minimal CRE promoter on the CREB binding and the reporter EGFP gene expression. ( A ) Artificial CRE-uno promoter: transcription start (broken arrow), CRE sequence (bold), AatII site (underlined), Nb.BsrDI nicking positions (vertical arrows), and positions of 5-mC/5-hmC/5-fC/5-caC in the incorporated synthetic oligonucleotides (asterisks). ( B ) Inhibition of the AatII restriction endonuclease by the M.SssI methylation of the central CG-dinucleotide in CRE. Agarose gel electrophoresis of pCRE-uno incubated with AatII. Arrows indicate migration positions of the linearized vector (4408 bp) and of the covalently closed (cc) and nicked (nc) forms of circular vector DNA. ( C ) Verification of the incorporation of synthetic oligonucleotides containing one or three 5-mC (1 × 5mC, 3 × 5mC) by inhibition of the AatII cleavage. ( D ) Scheme of the PciI/BmtI digest of the pCRE-uno vector and electrophoretic mobility shift assay (EMSA) of the PciI/BmtI fragments under the indicated CREB:DNA molar ratios. The shorter (180 bp) fragment contains CRE with no or one 5-mC in the central CG-dinucleotide. The right panel shows control CRE-less vector (pCRE-zero) incubated with CREB in parallel. ( E ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one (in the central CG-dinucleotide of CRE) or three 5mC. Representative FACS data and relative EGFP expression values (mean ± SD, n = 7). Expression in the absence of an EGFP-coding vector (DsRed) denotes the lower detection limit. CRE-less vector (pCRE-zero) was included in two experiments as a reference for the basal expression level. P -values calculated by the Student's t -test.

    Techniques Used: Methylation, Binding Assay, Expressing, Sequencing, Inhibition, Agarose Gel Electrophoresis, Incubation, Migration, Plasmid Preparation, Electrophoretic Mobility Shift Assay, Transfection, FACS

    Impacts of one or three 5-hmC in the minimal CRE promoter on the EGFP gene expression and comparison with the respective effects of 5-mC. ( A ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one or three 5-hmC. Representative fluorescence scatter plots and the correspondent overlaid fluorescence distribution plots of HeLa cells 24 h after transfection (on the left) as well as mean EGFP expression values determined relative to the construct containing unmodified oligonucleotide (‘C’). Mean of nine independent experiments ± SD; P -values calculated by the Student's t -test. ( B ) CREB binding to CRE containing one 5-mC or 5-hmC in the central CG-dinucleotide detected by EMSA of the pCRE-uno PciI/BmtI fragment. ( C ) EGFP expression in HeLa cells transfected with pCRE-uno containing either one (1×) or three (3×) of the indicated CpG modifications in the promoter fragment (relative to pCRE-uno without CpG modifications). Lines represent independent transfections.
    Figure Legend Snippet: Impacts of one or three 5-hmC in the minimal CRE promoter on the EGFP gene expression and comparison with the respective effects of 5-mC. ( A ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one or three 5-hmC. Representative fluorescence scatter plots and the correspondent overlaid fluorescence distribution plots of HeLa cells 24 h after transfection (on the left) as well as mean EGFP expression values determined relative to the construct containing unmodified oligonucleotide (‘C’). Mean of nine independent experiments ± SD; P -values calculated by the Student's t -test. ( B ) CREB binding to CRE containing one 5-mC or 5-hmC in the central CG-dinucleotide detected by EMSA of the pCRE-uno PciI/BmtI fragment. ( C ) EGFP expression in HeLa cells transfected with pCRE-uno containing either one (1×) or three (3×) of the indicated CpG modifications in the promoter fragment (relative to pCRE-uno without CpG modifications). Lines represent independent transfections.

    Techniques Used: Expressing, Transfection, Fluorescence, Construct, Binding Assay

    2) Product Images from "Multiple Acyl-CoA Dehydrogenation Deficiency (Glutaric Aciduria Type II) with a Novel Mutation of Electron Transfer Flavoprotein-Dehydrogenase in a Cat"

    Article Title: Multiple Acyl-CoA Dehydrogenation Deficiency (Glutaric Aciduria Type II) with a Novel Mutation of Electron Transfer Flavoprotein-Dehydrogenase in a Cat

    Journal: JIMD Reports

    doi: 10.1007/8904_2013_268

    The ETFDH mutation in the present cat with MADD. ( a ) The mutation c.692T > G ( arrow head ) was identified by sequencing analysis. ( b ) The allelic frequency of the mutation was determined by PCR-RFLP analysis. The PciI restriction site ( arrow ), ACATGT,
    Figure Legend Snippet: The ETFDH mutation in the present cat with MADD. ( a ) The mutation c.692T > G ( arrow head ) was identified by sequencing analysis. ( b ) The allelic frequency of the mutation was determined by PCR-RFLP analysis. The PciI restriction site ( arrow ), ACATGT,

    Techniques Used: Mutagenesis, Sequencing, Polymerase Chain Reaction

    Related Articles

    Amplification:

    Article Title: Multiple Acyl-CoA Dehydrogenation Deficiency (Glutaric Aciduria Type II) with a Novel Mutation of Electron Transfer Flavoprotein-Dehydrogenase in a Cat
    Article Snippet: .. The amplified ETFDH was reacted with the restriction enzyme PciI (New England Biolabs, Ipswich, MA, USA). ..

    In Vitro:

    Article Title: A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2
    Article Snippet: .. cDNA synthesis and RT fidelity assay The fidelity assays were performed as described previously ( ) with some modifications. lac Zα RNA for cDNA synthesis was produced in vitro by T7 RNA polymerase from New England Biolabs (NEB) and plasmid pBSC SK—linearized with PciI (NEB). .. The RNA was purified using Nucleospin RNA II-binding spin columns (Macherey–Nagel) and dissolved in RNase-free water.

    Electrophoretic Mobility Shift Assay:

    Article Title: Functional impacts of 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxycytosine at a single hemi-modified CpG dinucleotide in a gene promoter
    Article Snippet: .. Electrophoretic mobility shift assays Vectors pCRE-uno and pCRE-zero were digested with PciI and BmtI (both NEB) to produce the CRE-less 4228 bp fragment and the 180 bp fragment with one (pCRE-uno) or no CRE (pCRE-zero) and cleaned up. .. Binding reactions contained 10 nM DNA (400 ng in 15 μl reaction volume) and proportional variable amounts of the purified CREB protein (BioCat, Heidelberg, Germany) in the binding buffer composed of 20 mM Tris-HCl (pH 8.0), 50 mM KCl, 5 mM MgCl2 , 1 mM dithiothreitol and 5% glycerol (all reagents from Sigma-Aldrich, Seelze, Germany) supplemented with 1 mg/ml bovine serum albumin (NEB).

    Produced:

    Article Title: A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2
    Article Snippet: .. cDNA synthesis and RT fidelity assay The fidelity assays were performed as described previously ( ) with some modifications. lac Zα RNA for cDNA synthesis was produced in vitro by T7 RNA polymerase from New England Biolabs (NEB) and plasmid pBSC SK—linearized with PciI (NEB). .. The RNA was purified using Nucleospin RNA II-binding spin columns (Macherey–Nagel) and dissolved in RNase-free water.

    Derivative Assay:

    Article Title: Construction of a thermo-sensitive pRI857 vector for efficient DNA capturing in Escherichia coli.
    Article Snippet: .. To establish a positive cloning system with a zero background for high-throughput DNA cloning purpose. .. To establish a positive cloning system with a zero background for high-throughput DNA cloning purpose.

    Expressing:

    Article Title: The Moving Junction Protein RON8 Facilitates Firm Attachment and Host Cell Invasion in Toxoplasma gondii
    Article Snippet: .. The 12 kb RON8-HA expression cassette was subcloned into this vector at an EcoRV site following excision from pGRA-HA_HPT-RON8 by PciI and DraIII digestion and blunting with Klenow fragment (NEB). .. Sequencing established the inverse orientation of the cassette relative to the KU80 flanks and confirmed placement of the HPT and RON8-HA cassettes between these flanks.

    Polymerase Chain Reaction:

    Article Title: Gene Therapy for Human Lung Adenocarcinoma Using a Suicide Gene Driven by a Lung-Specific Promoter Delivered by JC Virus-Like Particles
    Article Snippet: .. The PCR product was digested with PciI and SalI (NEB, Ipswich, MA) and ligated using T4 DNA ligase (NEB) with pEGFP-N1 plasmid DNA (Clontech Laboratories, Inc., Mountain View, CA) from which the CMV promoter had been removed by digestion with PciI and SalI. .. The resulting plasmid, pSPB-gfp, was confirmed by DNA sequencing.

    Plasmid Preparation:

    Article Title: A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2
    Article Snippet: .. cDNA synthesis and RT fidelity assay The fidelity assays were performed as described previously ( ) with some modifications. lac Zα RNA for cDNA synthesis was produced in vitro by T7 RNA polymerase from New England Biolabs (NEB) and plasmid pBSC SK—linearized with PciI (NEB). .. The RNA was purified using Nucleospin RNA II-binding spin columns (Macherey–Nagel) and dissolved in RNase-free water.

    Article Title: Gene Therapy for Human Lung Adenocarcinoma Using a Suicide Gene Driven by a Lung-Specific Promoter Delivered by JC Virus-Like Particles
    Article Snippet: .. The PCR product was digested with PciI and SalI (NEB, Ipswich, MA) and ligated using T4 DNA ligase (NEB) with pEGFP-N1 plasmid DNA (Clontech Laboratories, Inc., Mountain View, CA) from which the CMV promoter had been removed by digestion with PciI and SalI. .. The resulting plasmid, pSPB-gfp, was confirmed by DNA sequencing.

    Article Title: Construction of a thermo-sensitive pRI857 vector for efficient DNA capturing in Escherichia coli.
    Article Snippet: .. To establish a positive cloning system with a zero background for high-throughput DNA cloning purpose. .. To establish a positive cloning system with a zero background for high-throughput DNA cloning purpose.

    Article Title: Anticooperative Binding Governs the Mechanics of Ethidium-Complexed DNA
    Article Snippet: .. DNA constructA 10,929 bp linear DNA fragment was excised from a custom-made 11.3 kbp plasmid by digestion with the restriction enzymes PciI and SacI. .. Biotin- or digoxigenin-modified attachment handles were produced by digesting 1.2 kbp biotin- and a digoxigenin dUTP-labeled PCR fragments ( ) from plasmid pBluescript II SK+ (Stratagene, La Jolla, CA) with PciI and SacI (New England Biolabs, Ipswich, MA), respectively, approximately in the middle of the fragments.

    Article Title: The Moving Junction Protein RON8 Facilitates Firm Attachment and Host Cell Invasion in Toxoplasma gondii
    Article Snippet: .. The 12 kb RON8-HA expression cassette was subcloned into this vector at an EcoRV site following excision from pGRA-HA_HPT-RON8 by PciI and DraIII digestion and blunting with Klenow fragment (NEB). .. Sequencing established the inverse orientation of the cassette relative to the KU80 flanks and confirmed placement of the HPT and RON8-HA cassettes between these flanks.

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  • 94
    New England Biolabs pcii
    Impact of CpG hemi-methylation in the minimal CRE promoter on the CREB binding and the reporter EGFP gene expression. ( A ) Artificial CRE-uno promoter: transcription start (broken arrow), CRE sequence (bold), AatII site (underlined), Nb.BsrDI nicking positions (vertical arrows), and positions of 5-mC/5-hmC/5-fC/5-caC in the incorporated synthetic oligonucleotides (asterisks). ( B ) Inhibition of the AatII restriction endonuclease by the M.SssI methylation of the central CG-dinucleotide in CRE. Agarose gel electrophoresis of <t>pCRE-uno</t> incubated with AatII. Arrows indicate migration positions of the linearized vector (4408 bp) and of the covalently closed (cc) and nicked (nc) forms of circular vector DNA. ( C ) Verification of the incorporation of synthetic oligonucleotides containing one or three 5-mC (1 × 5mC, 3 × 5mC) by inhibition of the AatII cleavage. ( D ) Scheme of the <t>PciI/BmtI</t> digest of the pCRE-uno vector and electrophoretic mobility shift assay (EMSA) of the PciI/BmtI fragments under the indicated CREB:DNA molar ratios. The shorter (180 bp) fragment contains CRE with no or one 5-mC in the central CG-dinucleotide. The right panel shows control CRE-less vector (pCRE-zero) incubated with CREB in parallel. ( E ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one (in the central CG-dinucleotide of CRE) or three 5mC. Representative FACS data and relative EGFP expression values (mean ± SD, n = 7). Expression in the absence of an EGFP-coding vector (DsRed) denotes the lower detection limit. CRE-less vector (pCRE-zero) was included in two experiments as a reference for the basal expression level. P -values calculated by the Student's t -test.
    Pcii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcii/product/New England Biolabs
    Average 94 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    pcii - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    Image Search Results


    Impact of CpG hemi-methylation in the minimal CRE promoter on the CREB binding and the reporter EGFP gene expression. ( A ) Artificial CRE-uno promoter: transcription start (broken arrow), CRE sequence (bold), AatII site (underlined), Nb.BsrDI nicking positions (vertical arrows), and positions of 5-mC/5-hmC/5-fC/5-caC in the incorporated synthetic oligonucleotides (asterisks). ( B ) Inhibition of the AatII restriction endonuclease by the M.SssI methylation of the central CG-dinucleotide in CRE. Agarose gel electrophoresis of pCRE-uno incubated with AatII. Arrows indicate migration positions of the linearized vector (4408 bp) and of the covalently closed (cc) and nicked (nc) forms of circular vector DNA. ( C ) Verification of the incorporation of synthetic oligonucleotides containing one or three 5-mC (1 × 5mC, 3 × 5mC) by inhibition of the AatII cleavage. ( D ) Scheme of the PciI/BmtI digest of the pCRE-uno vector and electrophoretic mobility shift assay (EMSA) of the PciI/BmtI fragments under the indicated CREB:DNA molar ratios. The shorter (180 bp) fragment contains CRE with no or one 5-mC in the central CG-dinucleotide. The right panel shows control CRE-less vector (pCRE-zero) incubated with CREB in parallel. ( E ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one (in the central CG-dinucleotide of CRE) or three 5mC. Representative FACS data and relative EGFP expression values (mean ± SD, n = 7). Expression in the absence of an EGFP-coding vector (DsRed) denotes the lower detection limit. CRE-less vector (pCRE-zero) was included in two experiments as a reference for the basal expression level. P -values calculated by the Student's t -test.

    Journal: Nucleic Acids Research

    Article Title: Functional impacts of 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxycytosine at a single hemi-modified CpG dinucleotide in a gene promoter

    doi: 10.1093/nar/gkx718

    Figure Lengend Snippet: Impact of CpG hemi-methylation in the minimal CRE promoter on the CREB binding and the reporter EGFP gene expression. ( A ) Artificial CRE-uno promoter: transcription start (broken arrow), CRE sequence (bold), AatII site (underlined), Nb.BsrDI nicking positions (vertical arrows), and positions of 5-mC/5-hmC/5-fC/5-caC in the incorporated synthetic oligonucleotides (asterisks). ( B ) Inhibition of the AatII restriction endonuclease by the M.SssI methylation of the central CG-dinucleotide in CRE. Agarose gel electrophoresis of pCRE-uno incubated with AatII. Arrows indicate migration positions of the linearized vector (4408 bp) and of the covalently closed (cc) and nicked (nc) forms of circular vector DNA. ( C ) Verification of the incorporation of synthetic oligonucleotides containing one or three 5-mC (1 × 5mC, 3 × 5mC) by inhibition of the AatII cleavage. ( D ) Scheme of the PciI/BmtI digest of the pCRE-uno vector and electrophoretic mobility shift assay (EMSA) of the PciI/BmtI fragments under the indicated CREB:DNA molar ratios. The shorter (180 bp) fragment contains CRE with no or one 5-mC in the central CG-dinucleotide. The right panel shows control CRE-less vector (pCRE-zero) incubated with CREB in parallel. ( E ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one (in the central CG-dinucleotide of CRE) or three 5mC. Representative FACS data and relative EGFP expression values (mean ± SD, n = 7). Expression in the absence of an EGFP-coding vector (DsRed) denotes the lower detection limit. CRE-less vector (pCRE-zero) was included in two experiments as a reference for the basal expression level. P -values calculated by the Student's t -test.

    Article Snippet: Electrophoretic mobility shift assays Vectors pCRE-uno and pCRE-zero were digested with PciI and BmtI (both NEB) to produce the CRE-less 4228 bp fragment and the 180 bp fragment with one (pCRE-uno) or no CRE (pCRE-zero) and cleaned up.

    Techniques: Methylation, Binding Assay, Expressing, Sequencing, Inhibition, Agarose Gel Electrophoresis, Incubation, Migration, Plasmid Preparation, Electrophoretic Mobility Shift Assay, Transfection, FACS

    Impacts of one or three 5-hmC in the minimal CRE promoter on the EGFP gene expression and comparison with the respective effects of 5-mC. ( A ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one or three 5-hmC. Representative fluorescence scatter plots and the correspondent overlaid fluorescence distribution plots of HeLa cells 24 h after transfection (on the left) as well as mean EGFP expression values determined relative to the construct containing unmodified oligonucleotide (‘C’). Mean of nine independent experiments ± SD; P -values calculated by the Student's t -test. ( B ) CREB binding to CRE containing one 5-mC or 5-hmC in the central CG-dinucleotide detected by EMSA of the pCRE-uno PciI/BmtI fragment. ( C ) EGFP expression in HeLa cells transfected with pCRE-uno containing either one (1×) or three (3×) of the indicated CpG modifications in the promoter fragment (relative to pCRE-uno without CpG modifications). Lines represent independent transfections.

    Journal: Nucleic Acids Research

    Article Title: Functional impacts of 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxycytosine at a single hemi-modified CpG dinucleotide in a gene promoter

    doi: 10.1093/nar/gkx718

    Figure Lengend Snippet: Impacts of one or three 5-hmC in the minimal CRE promoter on the EGFP gene expression and comparison with the respective effects of 5-mC. ( A ) EGFP expression in HeLa cells transfected with pCRE-uno containing none, one or three 5-hmC. Representative fluorescence scatter plots and the correspondent overlaid fluorescence distribution plots of HeLa cells 24 h after transfection (on the left) as well as mean EGFP expression values determined relative to the construct containing unmodified oligonucleotide (‘C’). Mean of nine independent experiments ± SD; P -values calculated by the Student's t -test. ( B ) CREB binding to CRE containing one 5-mC or 5-hmC in the central CG-dinucleotide detected by EMSA of the pCRE-uno PciI/BmtI fragment. ( C ) EGFP expression in HeLa cells transfected with pCRE-uno containing either one (1×) or three (3×) of the indicated CpG modifications in the promoter fragment (relative to pCRE-uno without CpG modifications). Lines represent independent transfections.

    Article Snippet: Electrophoretic mobility shift assays Vectors pCRE-uno and pCRE-zero were digested with PciI and BmtI (both NEB) to produce the CRE-less 4228 bp fragment and the 180 bp fragment with one (pCRE-uno) or no CRE (pCRE-zero) and cleaned up.

    Techniques: Expressing, Transfection, Fluorescence, Construct, Binding Assay

    The ETFDH mutation in the present cat with MADD. ( a ) The mutation c.692T > G ( arrow head ) was identified by sequencing analysis. ( b ) The allelic frequency of the mutation was determined by PCR-RFLP analysis. The PciI restriction site ( arrow ), ACATGT,

    Journal: JIMD Reports

    Article Title: Multiple Acyl-CoA Dehydrogenation Deficiency (Glutaric Aciduria Type II) with a Novel Mutation of Electron Transfer Flavoprotein-Dehydrogenase in a Cat

    doi: 10.1007/8904_2013_268

    Figure Lengend Snippet: The ETFDH mutation in the present cat with MADD. ( a ) The mutation c.692T > G ( arrow head ) was identified by sequencing analysis. ( b ) The allelic frequency of the mutation was determined by PCR-RFLP analysis. The PciI restriction site ( arrow ), ACATGT,

    Article Snippet: The amplified ETFDH was reacted with the restriction enzyme PciI (New England Biolabs, Ipswich, MA, USA).

    Techniques: Mutagenesis, Sequencing, Polymerase Chain Reaction