klenow dna polymerase  (Thermo Fisher)


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    Name:
    DNA Polymerase I Large Klenow Fragment
    Description:
    DNA Polymerase I Large Klenow Fragment is a DNA polymerase enzyme that lacks the 5 to 3 exonuclease activity of intact DNA Polymerase I but does exhibit the 5 to 3 DNA polymerase and 3 to 5 exonuclease activities Applications Fill in of 5 overhangs 1 Synthesis of probes by random primers labeling method 2 Sequencing single and double stranded DNA 3 Site directed mutagenesis Source Purified from E coli expressing the Klenow fragment on a plasmid Performance and Quality Testing SDS PAGE purity single stranded and double stranded endodeoxyribonuclease and self priming assays performance evaluated in fill in reaction Unit Definition One unit incorporates 10 nmol of total deoxyribonucleotide into acid precipitable material in 30 min at 37°C using a template•primer
    Catalog Number:
    18012021
    Price:
    None
    Applications:
    Cloning|Genotyping & Genomic Profiling|Restriction Enzyme Cloning|Gene Expression Analysis & Genotyping|Array CGH
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher klenow dna polymerase
    DNA Polymerase I Large Klenow Fragment is a DNA polymerase enzyme that lacks the 5 to 3 exonuclease activity of intact DNA Polymerase I but does exhibit the 5 to 3 DNA polymerase and 3 to 5 exonuclease activities Applications Fill in of 5 overhangs 1 Synthesis of probes by random primers labeling method 2 Sequencing single and double stranded DNA 3 Site directed mutagenesis Source Purified from E coli expressing the Klenow fragment on a plasmid Performance and Quality Testing SDS PAGE purity single stranded and double stranded endodeoxyribonuclease and self priming assays performance evaluated in fill in reaction Unit Definition One unit incorporates 10 nmol of total deoxyribonucleotide into acid precipitable material in 30 min at 37°C using a template•primer
    https://www.bioz.com/result/klenow dna polymerase/product/Thermo Fisher
    Average 93 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    klenow dna polymerase - by Bioz Stars, 2020-07
    93/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: A bacterial group II intron encoding reverse transcriptase, maturase, and DNA endonuclease activities: biochemical demonstration of maturase activity and insertion of new genetic information within the intron
    Article Snippet: .. To ensure that no unintended changes were introduced by PCR, the 2.4-kb Ssp I fragment was replaced with that of pLI1, and the remaining region was sequenced as described above for pLI1P. pLI1–FS is a derivative of pLI1, which has a frameshift after codon 124 of the LtrA ORF, introduced by linearizing pLI1 at the Age I site in the LtrA ORF and religating after filling in the 4-nucleotide 5′ overhangs with Klenow DNA polymerase (Life Technologies). pLI1–DD− and pLI1–EBS1-6C are derivatives of pLI1 that were constructed by PCR mutagenesis. pLI1–DD− has the conserved YADD sequence in the RT domain changed to YAAA, and pLI1–EBS1-6C has a single nucleotide change in EBS1 (G284C) and a complementary change in IBS1 (E1–6G) to permit splicing. .. In each case, the region generated by PCR was sequenced completely to insure that no adventitious mutations had been introduced.

    Mutagenesis:

    Article Title: A bacterial group II intron encoding reverse transcriptase, maturase, and DNA endonuclease activities: biochemical demonstration of maturase activity and insertion of new genetic information within the intron
    Article Snippet: .. To ensure that no unintended changes were introduced by PCR, the 2.4-kb Ssp I fragment was replaced with that of pLI1, and the remaining region was sequenced as described above for pLI1P. pLI1–FS is a derivative of pLI1, which has a frameshift after codon 124 of the LtrA ORF, introduced by linearizing pLI1 at the Age I site in the LtrA ORF and religating after filling in the 4-nucleotide 5′ overhangs with Klenow DNA polymerase (Life Technologies). pLI1–DD− and pLI1–EBS1-6C are derivatives of pLI1 that were constructed by PCR mutagenesis. pLI1–DD− has the conserved YADD sequence in the RT domain changed to YAAA, and pLI1–EBS1-6C has a single nucleotide change in EBS1 (G284C) and a complementary change in IBS1 (E1–6G) to permit splicing. .. In each case, the region generated by PCR was sequenced completely to insure that no adventitious mutations had been introduced.

    Sequencing:

    Article Title: A bacterial group II intron encoding reverse transcriptase, maturase, and DNA endonuclease activities: biochemical demonstration of maturase activity and insertion of new genetic information within the intron
    Article Snippet: .. To ensure that no unintended changes were introduced by PCR, the 2.4-kb Ssp I fragment was replaced with that of pLI1, and the remaining region was sequenced as described above for pLI1P. pLI1–FS is a derivative of pLI1, which has a frameshift after codon 124 of the LtrA ORF, introduced by linearizing pLI1 at the Age I site in the LtrA ORF and religating after filling in the 4-nucleotide 5′ overhangs with Klenow DNA polymerase (Life Technologies). pLI1–DD− and pLI1–EBS1-6C are derivatives of pLI1 that were constructed by PCR mutagenesis. pLI1–DD− has the conserved YADD sequence in the RT domain changed to YAAA, and pLI1–EBS1-6C has a single nucleotide change in EBS1 (G284C) and a complementary change in IBS1 (E1–6G) to permit splicing. .. In each case, the region generated by PCR was sequenced completely to insure that no adventitious mutations had been introduced.

    Construct:

    Article Title: A bacterial group II intron encoding reverse transcriptase, maturase, and DNA endonuclease activities: biochemical demonstration of maturase activity and insertion of new genetic information within the intron
    Article Snippet: .. To ensure that no unintended changes were introduced by PCR, the 2.4-kb Ssp I fragment was replaced with that of pLI1, and the remaining region was sequenced as described above for pLI1P. pLI1–FS is a derivative of pLI1, which has a frameshift after codon 124 of the LtrA ORF, introduced by linearizing pLI1 at the Age I site in the LtrA ORF and religating after filling in the 4-nucleotide 5′ overhangs with Klenow DNA polymerase (Life Technologies). pLI1–DD− and pLI1–EBS1-6C are derivatives of pLI1 that were constructed by PCR mutagenesis. pLI1–DD− has the conserved YADD sequence in the RT domain changed to YAAA, and pLI1–EBS1-6C has a single nucleotide change in EBS1 (G284C) and a complementary change in IBS1 (E1–6G) to permit splicing. .. In each case, the region generated by PCR was sequenced completely to insure that no adventitious mutations had been introduced.

    Staining:

    Article Title: Effect of minichromosome maintenance protein 2 deficiency on the locations of DNA replication origins
    Article Snippet: .. For determination of yield, RNase I released material was converted to double-stranded DNA by random priming and treatment with the Klenow fragment of E. coli DNA polymerase I (Invitrogen 18012-021) and quantified using pico-green staining. ..

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  • 99
    Thermo Fisher dna polymerase i
    Evaluation of the HexaPrime assay. (A) Evaluation of different primer pairs for the detection of coronaviruses. Analysis was conducted using the HCoV-NL63 virus and all primer sets given in Table 2 were tested. Only amplification with primer sets 2, 4, 5 and 8 yielded distinct bands. Sequencing of products and analysis of fragment size revealed that only primer set 2 allowed efficient amplification of the desired product. M: size marker; mock-infected (−) or HCoV-NL63-infected (+) cell culture supernatant. (B) Detection of HCoV-NL63 and HCoV-HKU1 with the HexaPrime assay using primer set 2. All experimental procedures were conducted as described in Section 2 . M: size marker; W: water; NL63 and HKU1: mock-infected (−) or virus-infected (+) cell culture supernatant. (C) Sensitivity of the HexaPrime assay. Concentrated samples containing viral RNA (10 9 copies ml −1 ) were subjected to 10-fold serial dilutions in cell culture supernatant and the HexaPrime assay was conducted. For each RNA concentration, three different enzymes for SS <t>DNA</t> synthesis were trialed. A, B and C denote DNA Polymerase I, T7 Polymerase, and Sequenase 2.0, respectively.
    Dna Polymerase I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna polymerase i/product/Thermo Fisher
    Average 99 stars, based on 62 article reviews
    Price from $9.99 to $1999.99
    dna polymerase i - by Bioz Stars, 2020-07
    99/100 stars
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    Evaluation of the HexaPrime assay. (A) Evaluation of different primer pairs for the detection of coronaviruses. Analysis was conducted using the HCoV-NL63 virus and all primer sets given in Table 2 were tested. Only amplification with primer sets 2, 4, 5 and 8 yielded distinct bands. Sequencing of products and analysis of fragment size revealed that only primer set 2 allowed efficient amplification of the desired product. M: size marker; mock-infected (−) or HCoV-NL63-infected (+) cell culture supernatant. (B) Detection of HCoV-NL63 and HCoV-HKU1 with the HexaPrime assay using primer set 2. All experimental procedures were conducted as described in Section 2 . M: size marker; W: water; NL63 and HKU1: mock-infected (−) or virus-infected (+) cell culture supernatant. (C) Sensitivity of the HexaPrime assay. Concentrated samples containing viral RNA (10 9 copies ml −1 ) were subjected to 10-fold serial dilutions in cell culture supernatant and the HexaPrime assay was conducted. For each RNA concentration, three different enzymes for SS DNA synthesis were trialed. A, B and C denote DNA Polymerase I, T7 Polymerase, and Sequenase 2.0, respectively.

    Journal: Journal of Virological Methods

    Article Title: HexaPrime: A novel method for detection of coronaviruses

    doi: 10.1016/j.jviromet.2012.11.039

    Figure Lengend Snippet: Evaluation of the HexaPrime assay. (A) Evaluation of different primer pairs for the detection of coronaviruses. Analysis was conducted using the HCoV-NL63 virus and all primer sets given in Table 2 were tested. Only amplification with primer sets 2, 4, 5 and 8 yielded distinct bands. Sequencing of products and analysis of fragment size revealed that only primer set 2 allowed efficient amplification of the desired product. M: size marker; mock-infected (−) or HCoV-NL63-infected (+) cell culture supernatant. (B) Detection of HCoV-NL63 and HCoV-HKU1 with the HexaPrime assay using primer set 2. All experimental procedures were conducted as described in Section 2 . M: size marker; W: water; NL63 and HKU1: mock-infected (−) or virus-infected (+) cell culture supernatant. (C) Sensitivity of the HexaPrime assay. Concentrated samples containing viral RNA (10 9 copies ml −1 ) were subjected to 10-fold serial dilutions in cell culture supernatant and the HexaPrime assay was conducted. For each RNA concentration, three different enzymes for SS DNA synthesis were trialed. A, B and C denote DNA Polymerase I, T7 Polymerase, and Sequenase 2.0, respectively.

    Article Snippet: The efficiency of different SS synthesis enzymes, T7 Polymerase (Thermo Scientific, Vilnius, Lithuania), DNA Polymerase I (Thermo Scientific, Vilnius, Lithuania), and Sequenase 2.0 (Affymetrix, United Kingdom), was evaluated by means of densitometry following bands separation on a 1.5% agarose gel.

    Techniques: Amplification, Sequencing, Marker, Infection, Cell Culture, Concentration Assay, DNA Synthesis

    Schematic diagram of strand-specific library synthesis mechanism . mRNAs are fragmented by heat and magnesium (1) and primed for cDNA synthesis by an adapter-containing oligonucleotide (2,3) . Size selection and cleanup removes unincorperated oligonucleotides and small cDNA fragments (4) . Transient duplex breathing at the terminus of the RNA-cDNA hybrid (5) facilitates interaction with the single-stranded portion of the 5-prime capturing adapter (6) and E. coli DNA Polymerase I catalyses its incorporation into a complete library molecule (7) .

    Journal: Frontiers in Plant Science

    Article Title: BrAD-seq: Breath Adapter Directional sequencing: a streamlined, ultra-simple and fast library preparation protocol for strand specific mRNA library construction

    doi: 10.3389/fpls.2015.00366

    Figure Lengend Snippet: Schematic diagram of strand-specific library synthesis mechanism . mRNAs are fragmented by heat and magnesium (1) and primed for cDNA synthesis by an adapter-containing oligonucleotide (2,3) . Size selection and cleanup removes unincorperated oligonucleotides and small cDNA fragments (4) . Transient duplex breathing at the terminus of the RNA-cDNA hybrid (5) facilitates interaction with the single-stranded portion of the 5-prime capturing adapter (6) and E. coli DNA Polymerase I catalyses its incorporation into a complete library molecule (7) .

    Article Snippet: Subsequently, 6 μl of the following reaction mixture was added, mixed by pipetting to fully re-suspend the pellet and incubated at room temperature for 15 min: 3.5 μl H2 O, 1 μl 10X Thermo Pol I reaction buffer (Thermo Scientific, Cat. # EP0041), 1 μl 250 mM MgCl2 (made fresh and stored at −20 C), 0.25 μl 25 mM dNTPs (Thermo Scientific, Cat. # R1121), 0.25 μl Thermo DNA Pol I (Thermo Scientific, Cat. # EP0041) (10 μl total reaction volume).

    Techniques: Selection