expand high fidelity polymerase  (Thermo Fisher)


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    Name:
    Poly A Polymerase Yeast
    Description:
    Poly A Polymerase catalyses the addition of adenosine residues onto the 3 ends of RNA It can be used to add poly A tails to RNA in the first step of cloning The reaction requires Mn2 or Mg2 ATP as substrate and any RNA containing 3 hydroxyl termini as primer Substitution of cordycepin 5 triphosphate 3 dATP for ATP results in addition of a single 3 dA residue to the ends of the RNA a useful technique for labeling RNA at the 3 end In comparative studies yeast poly A polymerase works more efficiently than E coli poly A polymerase for RNA oligonucleotide labeling and poly A tailing Shorter incubation times are required for the yeast enzyme and it is found to label both long and short substrates equally well Poly A Polymerase is recommended over T4 RNA Ligase for 3 end labeling of long RNA molecules Note Various modified nucleotides can also be used to label the 3 end of RNA using Yeast Poly A Polymerase Purity Ribonuclease free Storage Buffer 20 mM Tris HCl pH 8 0 50 mM KCl 0 5 mM DTT 50 glycerol Assay Conditions 2 5mM Tris HCl pH 7 0 40 mM KCl 0 5 mM MnCl2 0 05 mM EDTA 0 5 mM DTT 0 2 mg mL BSA 10 glycerol 3 3 µM radiolabeled ATP 0 5mM ATP 6 5 µg poly A 100 bases poly A polymerase After incubation at 37°C for 10 min acid insoluble radioactivity is determined Unit Definition One unit equals 15 pmol min AMP incorporated into riboA 15 at 37°C Concentration 600 units µL Functional Test 3 end labeling of a ribonucleotide with cordycepin 5 triphosphate Functionally Tested 5X Poly A Polymerase Reaction Buffer 1 mL included PN 74226 100 mM Tris HCl pH 7 0 3 0 mM MnCl2 0 1 mM EDTA 1mM DTT 500 µg mL acetylated BSA 50 glycerol Applications 1 Addition of poly A tails to RNA 2 Labeling the 3 ends of RNA Source E coli strain containing an overproducing clone of Yeast Poly A Polymerase
    Catalog Number:
    74225Z25KU
    Price:
    None
    Category:
    Proteins Enzymes Peptides
    Applications:
    Nucleic Acid Labeling & Oligo Synthesis|PCR & Real-Time PCR|Reverse Transcription
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    Structured Review

    Thermo Fisher expand high fidelity polymerase
    Poly A Polymerase catalyses the addition of adenosine residues onto the 3 ends of RNA It can be used to add poly A tails to RNA in the first step of cloning The reaction requires Mn2 or Mg2 ATP as substrate and any RNA containing 3 hydroxyl termini as primer Substitution of cordycepin 5 triphosphate 3 dATP for ATP results in addition of a single 3 dA residue to the ends of the RNA a useful technique for labeling RNA at the 3 end In comparative studies yeast poly A polymerase works more efficiently than E coli poly A polymerase for RNA oligonucleotide labeling and poly A tailing Shorter incubation times are required for the yeast enzyme and it is found to label both long and short substrates equally well Poly A Polymerase is recommended over T4 RNA Ligase for 3 end labeling of long RNA molecules Note Various modified nucleotides can also be used to label the 3 end of RNA using Yeast Poly A Polymerase Purity Ribonuclease free Storage Buffer 20 mM Tris HCl pH 8 0 50 mM KCl 0 5 mM DTT 50 glycerol Assay Conditions 2 5mM Tris HCl pH 7 0 40 mM KCl 0 5 mM MnCl2 0 05 mM EDTA 0 5 mM DTT 0 2 mg mL BSA 10 glycerol 3 3 µM radiolabeled ATP 0 5mM ATP 6 5 µg poly A 100 bases poly A polymerase After incubation at 37°C for 10 min acid insoluble radioactivity is determined Unit Definition One unit equals 15 pmol min AMP incorporated into riboA 15 at 37°C Concentration 600 units µL Functional Test 3 end labeling of a ribonucleotide with cordycepin 5 triphosphate Functionally Tested 5X Poly A Polymerase Reaction Buffer 1 mL included PN 74226 100 mM Tris HCl pH 7 0 3 0 mM MnCl2 0 1 mM EDTA 1mM DTT 500 µg mL acetylated BSA 50 glycerol Applications 1 Addition of poly A tails to RNA 2 Labeling the 3 ends of RNA Source E coli strain containing an overproducing clone of Yeast Poly A Polymerase
    https://www.bioz.com/result/expand high fidelity polymerase/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    expand high fidelity polymerase - by Bioz Stars, 2021-09
    97/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: Molecular analysis of Pseudomonas aeruginosa strains isolated from cystic fibrosis patients
    Article Snippet: .. The PCR was optimized and performed in a total volume of 25 µl consisted of 20 ng of DNA, 1 × Taq polymerase reaction buffer (Invitrogen by Life Technologies, CA, USA), 1 U Taq polymerase (Invitrogen by Life Technologies, Waltham, USA), 1.5 mM of MgCl2 (for ms211 1.0 mM and for ms222 2.0 mM of MgCl2 were used, respectively), 200 µM of each deoxynucleotide, 6% dimethyl sulfoxide (DMSO) and 0.2 µM of each primer. ..

    Article Title: Distribution and natural infection status of synantrophic triatomines (Hemiptera: Reduviidae), vectors of Trypanosoma cruzi, reveals new epidemiological scenarios for chagas disease in the Highlands of Colombia
    Article Snippet: .. The PCR was performed in a final volume of 25 μL containing 40–50 ng of genomic DNA, 1X of buffer, 0.04 mM of dNTP, 1.5 mM MgCl2, 0.4 μM of each primer (TCZ1 and TCZ2), and 0.05 U of Taq polymerase (Invitrogen, California, USA). ..

    Article Title: Arabidopsis Mediator subunit 17 connects transcription with DNA repair after UV-B exposure
    Article Snippet: .. PCR was done using Pfu (Invitrogen) polymerase under the following conditions: 94°C for 5 min; 40 cycles of 94°C for 30 sec, 55°C for 20 sec and 72°C for 30 sec; and finally, one cycle at 70°C for 2 min. PCR product was purified from the gel, cloned in a pBluescript vector and sequenced. ..

    Article Title: Distribution and natural infection status of synantrophic triatomines (Hemiptera: Reduviidae), vectors of Trypanosoma cruzi, reveals new epidemiological scenarios for chagas disease in the Highlands of Colombia
    Article Snippet: .. The PCR was performed in a final volume of 25 μL containing 40–50 ng of genomic DNA, 1X of a buffer, 0.25 mM of dNTP, 2 mM MgCl2, 0.4 μM of each primer, and 0.05 U of Taq polymerase (Invitrogen, California, USA). ..

    Article Title: Rab40–Cullin5 complex regulates EPLIN and actin cytoskeleton dynamics during cell migration
    Article Snippet: .. PCR was performed using Taq polymerase (Invitrogen). ..

    Incubation:

    Article Title: Synthesis of modified nucleotide polymers by the poly(U) polymerase Cid1: Application to direct RNA sequencing on nanopores
    Article Snippet: .. Polynucleotide tailing with S. cerevisiae Poly(A) PolymeraseTo add a homopolymer to the 3’ ends of RNAs using Poly(A) Polymerase, Yeast (ThermoScientific 74225Y/Z) a reaction containing (1x Poly(A) Polymerase reaction buffer, 200fmol RNA, 0.5mM NTP) is incubated at 37°C for 30 minutes, then two volumes of Gel Loading Buffer II (Invitrogen: AM8546G) are immediately added to stop the reaction. ..

    Amplification:

    Article Title: Increased PD-1 Level in Severe Cervical Injury Is Associated With the Rare Programmed Cell Death 1 (PDCD1) rs36084323 A Allele in a Dominant Model
    Article Snippet: .. The amplification reaction was performed in a final volume of 20 μL containing 1x of polymerase buffer (Applied Biosystems), 0.5 mM MgCl2 , 2% DMSO, 200 μM dNTP’s, 1.0 μM of each primer, 1.0 unit of Ampli-Taq Gold (Applied Biosystems) and 80-200 ng of genomic DNA for the amplification of a 962 bp-PD1 fragment. ..

    other:

    Article Title: Effect of 12-weeks elastic band resistance training on MyomiRs and osteoporosis markers in elderly women with Osteosarcopenic obesity: a randomized controlled trial
    Article Snippet: Serum miRNA was then extracted using the mirVana PARIS Ambion kit, followed by real-time polymerase chain reaction (RT-PCR).

    Purification:

    Article Title: Arabidopsis Mediator subunit 17 connects transcription with DNA repair after UV-B exposure
    Article Snippet: .. PCR was done using Pfu (Invitrogen) polymerase under the following conditions: 94°C for 5 min; 40 cycles of 94°C for 30 sec, 55°C for 20 sec and 72°C for 30 sec; and finally, one cycle at 70°C for 2 min. PCR product was purified from the gel, cloned in a pBluescript vector and sequenced. ..

    Clone Assay:

    Article Title: Arabidopsis Mediator subunit 17 connects transcription with DNA repair after UV-B exposure
    Article Snippet: .. PCR was done using Pfu (Invitrogen) polymerase under the following conditions: 94°C for 5 min; 40 cycles of 94°C for 30 sec, 55°C for 20 sec and 72°C for 30 sec; and finally, one cycle at 70°C for 2 min. PCR product was purified from the gel, cloned in a pBluescript vector and sequenced. ..

    Plasmid Preparation:

    Article Title: Arabidopsis Mediator subunit 17 connects transcription with DNA repair after UV-B exposure
    Article Snippet: .. PCR was done using Pfu (Invitrogen) polymerase under the following conditions: 94°C for 5 min; 40 cycles of 94°C for 30 sec, 55°C for 20 sec and 72°C for 30 sec; and finally, one cycle at 70°C for 2 min. PCR product was purified from the gel, cloned in a pBluescript vector and sequenced. ..

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