exonuclease iii mung bean nuclease deletion kit  (Stratagene)

 
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    Stratagene exonuclease iii mung bean nuclease deletion kit
    Analysis of polysomal mRNA. Data from spermidine- and arginine-grown cultures are shown on the left and right, respectively, and the tops of the gradients are to the right. (A) A 254 profile of 10 to 40% sucrose gradients. (B) Northern blot analysis of <t>spe-1</t> mRNA from the aga strain (IC3). (C) Northern blot analysis of spe-1 mRNA from transformant DMH3, lacking the 5′-UTR sequences between the Afl <t>III</t> and Nru I sites. (D) Northern blot analysis of tub mRNA of strain IC3 shown in panel B ( tub mRNA from DMH3 was similar). Polysomal fractions are aligned below their approximate positions in the gradient.
    Exonuclease Iii Mung Bean Nuclease Deletion Kit, supplied by Stratagene, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/exonuclease iii mung bean nuclease deletion kit/product/Stratagene
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    exonuclease iii mung bean nuclease deletion kit - by Bioz Stars, 2020-11
    80/100 stars

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    1) Product Images from "Polyamine Regulation of Ornithine Decarboxylase Synthesis in Neurospora crassa"

    Article Title: Polyamine Regulation of Ornithine Decarboxylase Synthesis in Neurospora crassa

    Journal: Molecular and Cellular Biology

    doi:

    Analysis of polysomal mRNA. Data from spermidine- and arginine-grown cultures are shown on the left and right, respectively, and the tops of the gradients are to the right. (A) A 254 profile of 10 to 40% sucrose gradients. (B) Northern blot analysis of spe-1 mRNA from the aga strain (IC3). (C) Northern blot analysis of spe-1 mRNA from transformant DMH3, lacking the 5′-UTR sequences between the Afl III and Nru I sites. (D) Northern blot analysis of tub mRNA of strain IC3 shown in panel B ( tub mRNA from DMH3 was similar). Polysomal fractions are aligned below their approximate positions in the gradient.
    Figure Legend Snippet: Analysis of polysomal mRNA. Data from spermidine- and arginine-grown cultures are shown on the left and right, respectively, and the tops of the gradients are to the right. (A) A 254 profile of 10 to 40% sucrose gradients. (B) Northern blot analysis of spe-1 mRNA from the aga strain (IC3). (C) Northern blot analysis of spe-1 mRNA from transformant DMH3, lacking the 5′-UTR sequences between the Afl III and Nru I sites. (D) Northern blot analysis of tub mRNA of strain IC3 shown in panel B ( tub mRNA from DMH3 was similar). Polysomal fractions are aligned below their approximate positions in the gradient.

    Techniques Used: Northern Blot

    Restriction map of the spe-1 gene and flanking sequences found in plasmids pPHL1 and pPH1. The boxed area represents the transcribed region, beginning with the right-pointing arrow. The coding sequence, interrupted by one intron, is shown in black. Abbreviations: MCS, multiple-cloning site; B, Bgl II; V, Eco V; C, Cla I; R, Eco RI; P, Pst I; S, Sac I; A, Afl III; N, Nru I; K, Kpn I; Sa, Sal I; H, Hin dIII.
    Figure Legend Snippet: Restriction map of the spe-1 gene and flanking sequences found in plasmids pPHL1 and pPH1. The boxed area represents the transcribed region, beginning with the right-pointing arrow. The coding sequence, interrupted by one intron, is shown in black. Abbreviations: MCS, multiple-cloning site; B, Bgl II; V, Eco V; C, Cla I; R, Eco RI; P, Pst I; S, Sac I; A, Afl III; N, Nru I; K, Kpn I; Sa, Sal I; H, Hin dIII.

    Techniques Used: Sequencing, Clone Assay

    Role of the UAR on the regulation of spe-1 genes lacking the Afl III- Nru I segment of the 5′-UTR. (A) Schematic of spe-1 genes lacking 5′-UTR sequences in which the spe-1 UAR is either present (DMH3) or absent (DMH4). The positions of the Afl III and Nru I sites of the 5′-UTR are indicated. (B) Northern blot analysis of repressed (SPD) and derepressed (ARG) cultures of the transformants. Northern blots were hybridized with probes derived from spe-1 cDNA or the coding region of the tub gene.
    Figure Legend Snippet: Role of the UAR on the regulation of spe-1 genes lacking the Afl III- Nru I segment of the 5′-UTR. (A) Schematic of spe-1 genes lacking 5′-UTR sequences in which the spe-1 UAR is either present (DMH3) or absent (DMH4). The positions of the Afl III and Nru I sites of the 5′-UTR are indicated. (B) Northern blot analysis of repressed (SPD) and derepressed (ARG) cultures of the transformants. Northern blots were hybridized with probes derived from spe-1 cDNA or the coding region of the tub gene.

    Techniques Used: Northern Blot, Derivative Assay

    Expression of various spe-1 and chimeric transcripts driven by the β-tubulin ( tub ) promoter of N. crassa . (A) Schematic diagram of tub :: spe-1 genes introduced into strain IC54, with functional regions of each gene listed across the top. The spe-1 sequences are represented by open boxes; and tub sequences are represented by shaded boxes. The positions of the Afl III and Nru I sites in the spe-1 5′-UTR are shown. (B) Northern blots of 10 μg of total RNA from repressed (SPD) or derepressed (ARG) cultures, probed with spe-1 cDNA or the coding region of tub DNA. The relative abundance of tub :: spe-1 mRNA in each transformant, normalized to tub mRNA and relative to that in DMH43/SPD, is given below the panel.
    Figure Legend Snippet: Expression of various spe-1 and chimeric transcripts driven by the β-tubulin ( tub ) promoter of N. crassa . (A) Schematic diagram of tub :: spe-1 genes introduced into strain IC54, with functional regions of each gene listed across the top. The spe-1 sequences are represented by open boxes; and tub sequences are represented by shaded boxes. The positions of the Afl III and Nru I sites in the spe-1 5′-UTR are shown. (B) Northern blots of 10 μg of total RNA from repressed (SPD) or derepressed (ARG) cultures, probed with spe-1 cDNA or the coding region of tub DNA. The relative abundance of tub :: spe-1 mRNA in each transformant, normalized to tub mRNA and relative to that in DMH43/SPD, is given below the panel.

    Techniques Used: Expressing, Functional Assay, Northern Blot

    Effects of 5′-to-3′ deletions of the spe-1 upstream region on ODC activity and derepression of spe-1 mRNA. (A) Schematic representation of the wild-type (P2) and deleted spe-1 genes integrated at the his-3 locus of strain IC2794-5. Distances from the major transcription start site, indicated by the arrow, are given in base pairs. The relative positions of the Pst I (−1000), Sac I (−167), and Afl III (+97) sites are also shown. ODC activity (in units per milligram of protein) of the transformants grown with 1 mM spermidine (SPD) or 1 mM arginine (ARG) are given to the right. (B) Northern blots of total RNA (10 μg) from repressed (left) and derepressed (right) cultures of these transformants were probed with spe-1 cDNA ( spe-1 ) or a fragment of the β-tubulin gene ( tub ), the latter as a loading control. Lanes: 1, P2; 2, PΔ1; 3, PΔ2; 4, PΔ3; 5, PΔ4; 6, PΔ5; 7, S8; 8, PΔ6; 9, PΔ7. (C) Approximately 25 μg of total RNA from the derepressed transformants was analyzed by primer extension reactions with the MH12 primer to determine the 5′ ends of their spe-1 transcripts. The molecular size marker on the right is given in nucleotides. Lanes are labeled as in panel B.
    Figure Legend Snippet: Effects of 5′-to-3′ deletions of the spe-1 upstream region on ODC activity and derepression of spe-1 mRNA. (A) Schematic representation of the wild-type (P2) and deleted spe-1 genes integrated at the his-3 locus of strain IC2794-5. Distances from the major transcription start site, indicated by the arrow, are given in base pairs. The relative positions of the Pst I (−1000), Sac I (−167), and Afl III (+97) sites are also shown. ODC activity (in units per milligram of protein) of the transformants grown with 1 mM spermidine (SPD) or 1 mM arginine (ARG) are given to the right. (B) Northern blots of total RNA (10 μg) from repressed (left) and derepressed (right) cultures of these transformants were probed with spe-1 cDNA ( spe-1 ) or a fragment of the β-tubulin gene ( tub ), the latter as a loading control. Lanes: 1, P2; 2, PΔ1; 3, PΔ2; 4, PΔ3; 5, PΔ4; 6, PΔ5; 7, S8; 8, PΔ6; 9, PΔ7. (C) Approximately 25 μg of total RNA from the derepressed transformants was analyzed by primer extension reactions with the MH12 primer to determine the 5′ ends of their spe-1 transcripts. The molecular size marker on the right is given in nucleotides. Lanes are labeled as in panel B.

    Techniques Used: Activity Assay, Northern Blot, Marker, Labeling

    Related Articles

    Sequencing:

    Article Title: The First Step of Gibberellin Biosynthesis in Pumpkin Is Catalyzed by at Least Two Copalyl Diphosphate Synthases Encoded by Differentially Regulated Genes
    Article Snippet: .. For sequencing of the CmCPS1 cDNA clones obtained from the library, a series of unidirectional deletions was constructed from both ends using the Exonuclease III mung bean nuclease deletion kit (Stratagene). .. For each expression construct in pGEX-4T-3, at least five independent clones were analyzed for in vitro activity.

    Clone Assay:

    Article Title: The First Step of Gibberellin Biosynthesis in Pumpkin Is Catalyzed by at Least Two Copalyl Diphosphate Synthases Encoded by Differentially Regulated Genes
    Article Snippet: .. For sequencing of the CmCPS1 cDNA clones obtained from the library, a series of unidirectional deletions was constructed from both ends using the Exonuclease III mung bean nuclease deletion kit (Stratagene). .. For each expression construct in pGEX-4T-3, at least five independent clones were analyzed for in vitro activity.

    Generated:

    Article Title: Temporal Gene Regulation of the Channel Catfish Virus (Ictalurid Herpesvirus 1) †
    Article Snippet: .. Nested deletion subclones of the IE3C cDNA were generated by using an exonuclease III-mung bean nuclease deletion kit (Stratagene) according to the manufacturer’s instructions after digestion with Sac I and Eco RI, generating a unique 3′-overhang restriction site and a unique 5′ restriction site between the insert and the 3′ site chosen. .. These fragments were sequenced by using a Sequenase version 2.0 DNA sequencing kit (United States Biochemical, Cleveland, Ohio) with 1,000 Ci of [α-35 S]dATP (Amersham Co.) per mmol and T3 or T7 primers according to manufacturer’s instructions.

    Construct:

    Article Title: The First Step of Gibberellin Biosynthesis in Pumpkin Is Catalyzed by at Least Two Copalyl Diphosphate Synthases Encoded by Differentially Regulated Genes
    Article Snippet: .. For sequencing of the CmCPS1 cDNA clones obtained from the library, a series of unidirectional deletions was constructed from both ends using the Exonuclease III mung bean nuclease deletion kit (Stratagene). .. For each expression construct in pGEX-4T-3, at least five independent clones were analyzed for in vitro activity.

    Plasmid Preparation:

    Article Title: Polyamine Regulation of Ornithine Decarboxylase Synthesis in Neurospora crassa
    Article Snippet: .. This plasmid was used as starting material for 5′-to-3′ deletions from the spe-1 upstream Pst I site using an exonuclease III/mung bean nuclease deletion kit (Stratagene) as specified by the manufacturer. .. The Bgl II site in the multiple-cloning site of the pDE1 vector was used to linearize the plasmid and then filled in with α-thiophosphate nucleoside triphosphates.

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    Stratagene exonuclease iii mung bean nuclease deletion kit
    Analysis of polysomal mRNA. Data from spermidine- and arginine-grown cultures are shown on the left and right, respectively, and the tops of the gradients are to the right. (A) A 254 profile of 10 to 40% sucrose gradients. (B) Northern blot analysis of <t>spe-1</t> mRNA from the aga strain (IC3). (C) Northern blot analysis of spe-1 mRNA from transformant DMH3, lacking the 5′-UTR sequences between the Afl <t>III</t> and Nru I sites. (D) Northern blot analysis of tub mRNA of strain IC3 shown in panel B ( tub mRNA from DMH3 was similar). Polysomal fractions are aligned below their approximate positions in the gradient.
    Exonuclease Iii Mung Bean Nuclease Deletion Kit, supplied by Stratagene, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/exonuclease iii mung bean nuclease deletion kit/product/Stratagene
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    exonuclease iii mung bean nuclease deletion kit - by Bioz Stars, 2020-11
    80/100 stars
      Buy from Supplier

    Image Search Results


    Analysis of polysomal mRNA. Data from spermidine- and arginine-grown cultures are shown on the left and right, respectively, and the tops of the gradients are to the right. (A) A 254 profile of 10 to 40% sucrose gradients. (B) Northern blot analysis of spe-1 mRNA from the aga strain (IC3). (C) Northern blot analysis of spe-1 mRNA from transformant DMH3, lacking the 5′-UTR sequences between the Afl III and Nru I sites. (D) Northern blot analysis of tub mRNA of strain IC3 shown in panel B ( tub mRNA from DMH3 was similar). Polysomal fractions are aligned below their approximate positions in the gradient.

    Journal: Molecular and Cellular Biology

    Article Title: Polyamine Regulation of Ornithine Decarboxylase Synthesis in Neurospora crassa

    doi:

    Figure Lengend Snippet: Analysis of polysomal mRNA. Data from spermidine- and arginine-grown cultures are shown on the left and right, respectively, and the tops of the gradients are to the right. (A) A 254 profile of 10 to 40% sucrose gradients. (B) Northern blot analysis of spe-1 mRNA from the aga strain (IC3). (C) Northern blot analysis of spe-1 mRNA from transformant DMH3, lacking the 5′-UTR sequences between the Afl III and Nru I sites. (D) Northern blot analysis of tub mRNA of strain IC3 shown in panel B ( tub mRNA from DMH3 was similar). Polysomal fractions are aligned below their approximate positions in the gradient.

    Article Snippet: This plasmid was used as starting material for 5′-to-3′ deletions from the spe-1 upstream Pst I site using an exonuclease III/mung bean nuclease deletion kit (Stratagene) as specified by the manufacturer.

    Techniques: Northern Blot

    Restriction map of the spe-1 gene and flanking sequences found in plasmids pPHL1 and pPH1. The boxed area represents the transcribed region, beginning with the right-pointing arrow. The coding sequence, interrupted by one intron, is shown in black. Abbreviations: MCS, multiple-cloning site; B, Bgl II; V, Eco V; C, Cla I; R, Eco RI; P, Pst I; S, Sac I; A, Afl III; N, Nru I; K, Kpn I; Sa, Sal I; H, Hin dIII.

    Journal: Molecular and Cellular Biology

    Article Title: Polyamine Regulation of Ornithine Decarboxylase Synthesis in Neurospora crassa

    doi:

    Figure Lengend Snippet: Restriction map of the spe-1 gene and flanking sequences found in plasmids pPHL1 and pPH1. The boxed area represents the transcribed region, beginning with the right-pointing arrow. The coding sequence, interrupted by one intron, is shown in black. Abbreviations: MCS, multiple-cloning site; B, Bgl II; V, Eco V; C, Cla I; R, Eco RI; P, Pst I; S, Sac I; A, Afl III; N, Nru I; K, Kpn I; Sa, Sal I; H, Hin dIII.

    Article Snippet: This plasmid was used as starting material for 5′-to-3′ deletions from the spe-1 upstream Pst I site using an exonuclease III/mung bean nuclease deletion kit (Stratagene) as specified by the manufacturer.

    Techniques: Sequencing, Clone Assay

    Role of the UAR on the regulation of spe-1 genes lacking the Afl III- Nru I segment of the 5′-UTR. (A) Schematic of spe-1 genes lacking 5′-UTR sequences in which the spe-1 UAR is either present (DMH3) or absent (DMH4). The positions of the Afl III and Nru I sites of the 5′-UTR are indicated. (B) Northern blot analysis of repressed (SPD) and derepressed (ARG) cultures of the transformants. Northern blots were hybridized with probes derived from spe-1 cDNA or the coding region of the tub gene.

    Journal: Molecular and Cellular Biology

    Article Title: Polyamine Regulation of Ornithine Decarboxylase Synthesis in Neurospora crassa

    doi:

    Figure Lengend Snippet: Role of the UAR on the regulation of spe-1 genes lacking the Afl III- Nru I segment of the 5′-UTR. (A) Schematic of spe-1 genes lacking 5′-UTR sequences in which the spe-1 UAR is either present (DMH3) or absent (DMH4). The positions of the Afl III and Nru I sites of the 5′-UTR are indicated. (B) Northern blot analysis of repressed (SPD) and derepressed (ARG) cultures of the transformants. Northern blots were hybridized with probes derived from spe-1 cDNA or the coding region of the tub gene.

    Article Snippet: This plasmid was used as starting material for 5′-to-3′ deletions from the spe-1 upstream Pst I site using an exonuclease III/mung bean nuclease deletion kit (Stratagene) as specified by the manufacturer.

    Techniques: Northern Blot, Derivative Assay

    Expression of various spe-1 and chimeric transcripts driven by the β-tubulin ( tub ) promoter of N. crassa . (A) Schematic diagram of tub :: spe-1 genes introduced into strain IC54, with functional regions of each gene listed across the top. The spe-1 sequences are represented by open boxes; and tub sequences are represented by shaded boxes. The positions of the Afl III and Nru I sites in the spe-1 5′-UTR are shown. (B) Northern blots of 10 μg of total RNA from repressed (SPD) or derepressed (ARG) cultures, probed with spe-1 cDNA or the coding region of tub DNA. The relative abundance of tub :: spe-1 mRNA in each transformant, normalized to tub mRNA and relative to that in DMH43/SPD, is given below the panel.

    Journal: Molecular and Cellular Biology

    Article Title: Polyamine Regulation of Ornithine Decarboxylase Synthesis in Neurospora crassa

    doi:

    Figure Lengend Snippet: Expression of various spe-1 and chimeric transcripts driven by the β-tubulin ( tub ) promoter of N. crassa . (A) Schematic diagram of tub :: spe-1 genes introduced into strain IC54, with functional regions of each gene listed across the top. The spe-1 sequences are represented by open boxes; and tub sequences are represented by shaded boxes. The positions of the Afl III and Nru I sites in the spe-1 5′-UTR are shown. (B) Northern blots of 10 μg of total RNA from repressed (SPD) or derepressed (ARG) cultures, probed with spe-1 cDNA or the coding region of tub DNA. The relative abundance of tub :: spe-1 mRNA in each transformant, normalized to tub mRNA and relative to that in DMH43/SPD, is given below the panel.

    Article Snippet: This plasmid was used as starting material for 5′-to-3′ deletions from the spe-1 upstream Pst I site using an exonuclease III/mung bean nuclease deletion kit (Stratagene) as specified by the manufacturer.

    Techniques: Expressing, Functional Assay, Northern Blot

    Effects of 5′-to-3′ deletions of the spe-1 upstream region on ODC activity and derepression of spe-1 mRNA. (A) Schematic representation of the wild-type (P2) and deleted spe-1 genes integrated at the his-3 locus of strain IC2794-5. Distances from the major transcription start site, indicated by the arrow, are given in base pairs. The relative positions of the Pst I (−1000), Sac I (−167), and Afl III (+97) sites are also shown. ODC activity (in units per milligram of protein) of the transformants grown with 1 mM spermidine (SPD) or 1 mM arginine (ARG) are given to the right. (B) Northern blots of total RNA (10 μg) from repressed (left) and derepressed (right) cultures of these transformants were probed with spe-1 cDNA ( spe-1 ) or a fragment of the β-tubulin gene ( tub ), the latter as a loading control. Lanes: 1, P2; 2, PΔ1; 3, PΔ2; 4, PΔ3; 5, PΔ4; 6, PΔ5; 7, S8; 8, PΔ6; 9, PΔ7. (C) Approximately 25 μg of total RNA from the derepressed transformants was analyzed by primer extension reactions with the MH12 primer to determine the 5′ ends of their spe-1 transcripts. The molecular size marker on the right is given in nucleotides. Lanes are labeled as in panel B.

    Journal: Molecular and Cellular Biology

    Article Title: Polyamine Regulation of Ornithine Decarboxylase Synthesis in Neurospora crassa

    doi:

    Figure Lengend Snippet: Effects of 5′-to-3′ deletions of the spe-1 upstream region on ODC activity and derepression of spe-1 mRNA. (A) Schematic representation of the wild-type (P2) and deleted spe-1 genes integrated at the his-3 locus of strain IC2794-5. Distances from the major transcription start site, indicated by the arrow, are given in base pairs. The relative positions of the Pst I (−1000), Sac I (−167), and Afl III (+97) sites are also shown. ODC activity (in units per milligram of protein) of the transformants grown with 1 mM spermidine (SPD) or 1 mM arginine (ARG) are given to the right. (B) Northern blots of total RNA (10 μg) from repressed (left) and derepressed (right) cultures of these transformants were probed with spe-1 cDNA ( spe-1 ) or a fragment of the β-tubulin gene ( tub ), the latter as a loading control. Lanes: 1, P2; 2, PΔ1; 3, PΔ2; 4, PΔ3; 5, PΔ4; 6, PΔ5; 7, S8; 8, PΔ6; 9, PΔ7. (C) Approximately 25 μg of total RNA from the derepressed transformants was analyzed by primer extension reactions with the MH12 primer to determine the 5′ ends of their spe-1 transcripts. The molecular size marker on the right is given in nucleotides. Lanes are labeled as in panel B.

    Article Snippet: This plasmid was used as starting material for 5′-to-3′ deletions from the spe-1 upstream Pst I site using an exonuclease III/mung bean nuclease deletion kit (Stratagene) as specified by the manufacturer.

    Techniques: Activity Assay, Northern Blot, Marker, Labeling