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    New England Biolabs dna adenylation kit
    Activity of the MthRnl and the K246A, K246Q and K97A mutants in different steps of the ligation reaction. (A) pH-dependence of <t>self-adenylation</t> activity of enzymes (step 1) in reactions with radioactive ATP. The reactions were carried out as described in the Methods and the products were resolved by an SDS-PAGE. Autoradiographs of the gels are shown. (B) pH-dependence of adenylation of 5’-phosphorylated, 3’-amino-blocked ssDNA (pDNA17-NH 2 ) (step 2) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The reaction products were separated by 15% urea-PAGE and visualized with SYBR Gold stain. (C) pH-dependence of deadenylation/hydrolysis of 5’ pre-adenylated <t>DNA</t> (AppDNA17-NH 2 ) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The products were analyzed on 15% urea-PAGE and stained with SYBR Gold. Enzymes used in each step of ligation reactions are indicated on the left. Position of DNA donor substrates and products are indicated on the right. The pH of the buffer is indicated at the bottom. The control (c) indicates the reaction without enzyme.
    Dna Adenylation Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 118 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    New England Biolabs 3 adapter
    Activity of the MthRnl and the K246A, K246Q and K97A mutants in different steps of the ligation reaction. (A) pH-dependence of <t>self-adenylation</t> activity of enzymes (step 1) in reactions with radioactive ATP. The reactions were carried out as described in the Methods and the products were resolved by an SDS-PAGE. Autoradiographs of the gels are shown. (B) pH-dependence of adenylation of 5’-phosphorylated, 3’-amino-blocked ssDNA (pDNA17-NH 2 ) (step 2) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The reaction products were separated by 15% urea-PAGE and visualized with SYBR Gold stain. (C) pH-dependence of deadenylation/hydrolysis of 5’ pre-adenylated <t>DNA</t> (AppDNA17-NH 2 ) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The products were analyzed on 15% urea-PAGE and stained with SYBR Gold. Enzymes used in each step of ligation reactions are indicated on the left. Position of DNA donor substrates and products are indicated on the right. The pH of the buffer is indicated at the bottom. The control (c) indicates the reaction without enzyme.
    3 Adapter, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 119 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    New England Biolabs pre adenylated 3 linker
    Activity of the MthRnl and the K246A, K246Q and K97A mutants in different steps of the ligation reaction. (A) pH-dependence of <t>self-adenylation</t> activity of enzymes (step 1) in reactions with radioactive ATP. The reactions were carried out as described in the Methods and the products were resolved by an SDS-PAGE. Autoradiographs of the gels are shown. (B) pH-dependence of adenylation of 5’-phosphorylated, 3’-amino-blocked ssDNA (pDNA17-NH 2 ) (step 2) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The reaction products were separated by 15% urea-PAGE and visualized with SYBR Gold stain. (C) pH-dependence of deadenylation/hydrolysis of 5’ pre-adenylated <t>DNA</t> (AppDNA17-NH 2 ) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The products were analyzed on 15% urea-PAGE and stained with SYBR Gold. Enzymes used in each step of ligation reactions are indicated on the left. Position of DNA donor substrates and products are indicated on the right. The pH of the buffer is indicated at the bottom. The control (c) indicates the reaction without enzyme.
    Pre Adenylated 3 Linker, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 88/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Activity of the MthRnl and the K246A, K246Q and K97A mutants in different steps of the ligation reaction. (A) pH-dependence of self-adenylation activity of enzymes (step 1) in reactions with radioactive ATP. The reactions were carried out as described in the Methods and the products were resolved by an SDS-PAGE. Autoradiographs of the gels are shown. (B) pH-dependence of adenylation of 5’-phosphorylated, 3’-amino-blocked ssDNA (pDNA17-NH 2 ) (step 2) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The reaction products were separated by 15% urea-PAGE and visualized with SYBR Gold stain. (C) pH-dependence of deadenylation/hydrolysis of 5’ pre-adenylated DNA (AppDNA17-NH 2 ) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The products were analyzed on 15% urea-PAGE and stained with SYBR Gold. Enzymes used in each step of ligation reactions are indicated on the left. Position of DNA donor substrates and products are indicated on the right. The pH of the buffer is indicated at the bottom. The control (c) indicates the reaction without enzyme.

    Journal: BMC Molecular Biology

    Article Title: Structure-function analysis of Methanobacterium thermoautotrophicum RNA ligase - engineering a thermostable ATP independent enzyme

    doi: 10.1186/1471-2199-13-24

    Figure Lengend Snippet: Activity of the MthRnl and the K246A, K246Q and K97A mutants in different steps of the ligation reaction. (A) pH-dependence of self-adenylation activity of enzymes (step 1) in reactions with radioactive ATP. The reactions were carried out as described in the Methods and the products were resolved by an SDS-PAGE. Autoradiographs of the gels are shown. (B) pH-dependence of adenylation of 5’-phosphorylated, 3’-amino-blocked ssDNA (pDNA17-NH 2 ) (step 2) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The reaction products were separated by 15% urea-PAGE and visualized with SYBR Gold stain. (C) pH-dependence of deadenylation/hydrolysis of 5’ pre-adenylated DNA (AppDNA17-NH 2 ) with wild type and the mutants of MthRnl under reaction conditions described in the Methods. The products were analyzed on 15% urea-PAGE and stained with SYBR Gold. Enzymes used in each step of ligation reactions are indicated on the left. Position of DNA donor substrates and products are indicated on the right. The pH of the buffer is indicated at the bottom. The control (c) indicates the reaction without enzyme.

    Article Snippet: Preparative adenylation of DNA and RNA linkers (donor substrates) were performed using 5’-DNA Adenylation kit (NEB) [ ] Table .

    Techniques: Activity Assay, Ligation, SDS Page, Polyacrylamide Gel Electrophoresis, Staining