α 32 p atp  (Thermo Fisher)


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    Structured Review

    Thermo Fisher α 32 p atp
    ATPase activities of wt 1a and 1a helicase mutants. (A) ATPase assay. No protein (lane 1) or the indicated amounts of GST (lanes 2 to 4), GST-1aC (lanes 5 to 7), or GST-1a-K691A (K691A) were incubated with [α- 32 <t>P]ATP</t> at 37°C for 30 min.
    α 32 P Atp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/α 32 p atp/product/Thermo Fisher
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    α 32 p atp - by Bioz Stars, 2020-04
    93/100 stars

    Images

    1) Product Images from "Brome Mosaic Virus 1a Nucleoside Triphosphatase/Helicase Domain Plays Crucial Roles in Recruiting RNA Replication Templates"

    Article Title: Brome Mosaic Virus 1a Nucleoside Triphosphatase/Helicase Domain Plays Crucial Roles in Recruiting RNA Replication Templates

    Journal: Journal of Virology

    doi: 10.1128/JVI.79.21.13747-13758.2005

    ATPase activities of wt 1a and 1a helicase mutants. (A) ATPase assay. No protein (lane 1) or the indicated amounts of GST (lanes 2 to 4), GST-1aC (lanes 5 to 7), or GST-1a-K691A (K691A) were incubated with [α- 32 P]ATP at 37°C for 30 min.
    Figure Legend Snippet: ATPase activities of wt 1a and 1a helicase mutants. (A) ATPase assay. No protein (lane 1) or the indicated amounts of GST (lanes 2 to 4), GST-1aC (lanes 5 to 7), or GST-1a-K691A (K691A) were incubated with [α- 32 P]ATP at 37°C for 30 min.

    Techniques Used: ATPase Assay, Incubation

    2) Product Images from "Affinity Labeling of Hepatitis C Virus Replicase with a Nucleotide Analog: Identification of binding site"

    Article Title: Affinity Labeling of Hepatitis C Virus Replicase with a Nucleotide Analog: Identification of binding site

    Journal: Biochemistry

    doi: 10.1021/bi301098g

    (A) Effect of alanine substitution at position 38, 382 and 491 on the RdRp activity of HCV NS5B The RdRp activity of the WT and its mutant derivatives (1 μM) were determined and expressed as percent relative activity with respect to the WT enzyme. (B) Photocrosslinking of 32 P-labeled RNA template primer to the wild type enzyme and its mutant derivatives. The 5′-[ 32 P]-labeled 37-mer self-annealing RNA template primer was used as the template primer to crosslink with the WT enzyme and its mutant derivatives. The 32 P labeled 37-mer TP (200 nM) was incubated on ice with 1 μM of enzyme in a final volume of 50 μl. The mixture was UV irradiated and the crosslinked E-TP covalent complexes were resolved by SDS PAGE and visualized by PhosphorImaging. (C) UV mediated Photocrosslinking of 5′-α- 32 P-ATP to the wild type enzyme and its mutant derivatives in the absence and presence of template primer. The wild type enzyme and its mutant derivatives were photo-crosslinked with [α- 32 P]-ATP in the absence of template primer (lanes 1–3) or in the presence of rU 15. rA 10 template primer with 3′-deoxy terminated primer terminus (lanes 4–6). The crosslinked complex was resolved on SDS PAGE and visualized by PhosphorImaging.
    Figure Legend Snippet: (A) Effect of alanine substitution at position 38, 382 and 491 on the RdRp activity of HCV NS5B The RdRp activity of the WT and its mutant derivatives (1 μM) were determined and expressed as percent relative activity with respect to the WT enzyme. (B) Photocrosslinking of 32 P-labeled RNA template primer to the wild type enzyme and its mutant derivatives. The 5′-[ 32 P]-labeled 37-mer self-annealing RNA template primer was used as the template primer to crosslink with the WT enzyme and its mutant derivatives. The 32 P labeled 37-mer TP (200 nM) was incubated on ice with 1 μM of enzyme in a final volume of 50 μl. The mixture was UV irradiated and the crosslinked E-TP covalent complexes were resolved by SDS PAGE and visualized by PhosphorImaging. (C) UV mediated Photocrosslinking of 5′-α- 32 P-ATP to the wild type enzyme and its mutant derivatives in the absence and presence of template primer. The wild type enzyme and its mutant derivatives were photo-crosslinked with [α- 32 P]-ATP in the absence of template primer (lanes 1–3) or in the presence of rU 15. rA 10 template primer with 3′-deoxy terminated primer terminus (lanes 4–6). The crosslinked complex was resolved on SDS PAGE and visualized by PhosphorImaging.

    Techniques Used: Activity Assay, Mutagenesis, Labeling, Incubation, Irradiation, SDS Page

    3) Product Images from "Isomorphic Emissive GTP Surrogate Facilitates Initiation and Elongation of in Vitro Transcription Reactions"

    Article Title: Isomorphic Emissive GTP Surrogate Facilitates Initiation and Elongation of in Vitro Transcription Reactions

    Journal: Journal of the American Chemical Society

    doi: 10.1021/ja5039227

    Transcription reactions with template 3 in the presence of th GTP. (a) Small scale transcription using trace α- 32 P ATP. Lane 1: control transcription reaction in the absence of GTP or th GTP. Lane 2: control reaction in the presence of all natural NTPs. Lane 3: reaction in the presence of equimolar concentration of th GTP and GTP. Lane 4: reaction in the presence of th GTP. Incorporation efficiencies of th GTP transcripts are reported with respect to transcription in the presence of GTP. All reactions were performed in triplicate, and the errors reported are the standard deviations. (b) Large scale transcription reaction using template 3 with all natural NTPs (lane 1 and 1′) or ATP, UTP, CTP, and th GTP (lanes 2 and 2′, 78 ± 12% isolated yield) with UV light at 254 nm (on TLC plate) and 302 nm (PL). The reaction was resolved by gel electrophoresis on a denaturing 20% polyacrylamide gel.
    Figure Legend Snippet: Transcription reactions with template 3 in the presence of th GTP. (a) Small scale transcription using trace α- 32 P ATP. Lane 1: control transcription reaction in the absence of GTP or th GTP. Lane 2: control reaction in the presence of all natural NTPs. Lane 3: reaction in the presence of equimolar concentration of th GTP and GTP. Lane 4: reaction in the presence of th GTP. Incorporation efficiencies of th GTP transcripts are reported with respect to transcription in the presence of GTP. All reactions were performed in triplicate, and the errors reported are the standard deviations. (b) Large scale transcription reaction using template 3 with all natural NTPs (lane 1 and 1′) or ATP, UTP, CTP, and th GTP (lanes 2 and 2′, 78 ± 12% isolated yield) with UV light at 254 nm (on TLC plate) and 302 nm (PL). The reaction was resolved by gel electrophoresis on a denaturing 20% polyacrylamide gel.

    Techniques Used: Concentration Assay, Isolation, Thin Layer Chromatography, Nucleic Acid Electrophoresis

    Related Articles

    Amplification:

    Article Title: mRNA Decapping and 5′-3′ Decay Contribute to the Regulation of ABA Signaling in Arabidopsis thaliana
    Article Snippet: .. Northern blots were performed using random primed probes amplified from cDNA template with appropriate primers and radioactively labeled with DECAprimeTM II kit (Ambion) and [α-32 P]ATP (Hartmann Analytics), or oligoribonucleotide probes radioactively labeled with PNK (Thermo Scientific) and [γ-32 P]ATP (Hartmann Analytics). .. Membranes were hybridized overnight with radioactive probes in PerfectHyb buffer (Sigma), washed, analyzed with PhosphorImager Typhoon FLA 9000 (GE Healthcare) and quantified with ImageJ software (Molecular Dynamics). mRNA half-life measurement experiments were carried out as described (Souret et al., ).

    Article Title: Structure and Expression of a Dhurrinase (?-Glucosidase) from Sorghum 1
    Article Snippet: After PCR, the amplification products were electrophoresed through a low-melting agarose gel (1%), excised, and purified using the Magic PCR Prep system according to the procedure provided by the manufacturer (Promega). .. The purified PCR products (the 1446- and 627-bp fragments) were labeled with [α-32 P]ATP using random hexamers and the Klenow fragment of DNA polymerase (Ambion, Austin, TX) at 37°C for 1 h. The labeled fragments were used as a probe in northern- and Southern-blot analyses.

    Synthesized:

    Article Title: The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase
    Article Snippet: Unlabeled and biotin-, Cy3-, and Cy5-labeled oligonucleotides (see Table S1 in the supplemental material) were synthesized at Sangon BioTech (Shanghai, China). .. To radiolabel a 5′ biotin-labeled oligonucleotide (S14) (see Table S1), the DNA fragment was annealed to oligonucleotide S20 and filled in with [α-32 P]ATP using Klenow fragment (Thermo).

    Autoradiography:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. The samples were analyzed by electrophoresis on a 5% polyacrylamide (37.5:1 acrylamide/bisacrylamide) gel and then autoradiography.

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. The samples were analyzed by electrophoresis on a 5% polyacrylamide (37.5:1 acrylamide/bisacrylamide) gel followed by autoradiography.

    Electrophoresis:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. The samples were analyzed by electrophoresis on a 5% polyacrylamide (37.5:1 acrylamide/bisacrylamide) gel and then autoradiography.

    Article Title: The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase
    Article Snippet: To radiolabel a 5′ biotin-labeled oligonucleotide (S14) (see Table S1), the DNA fragment was annealed to oligonucleotide S20 and filled in with [α-32 P]ATP using Klenow fragment (Thermo). .. After the sample was boiled for 3 min and subsequently cooled on ice, it was subjected to electrophoresis in a 15% polyacrylamide gel containing 8 M urea in 1× Tris-borate-EDTA (TBE) buffer.

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. The samples were analyzed by electrophoresis on a 5% polyacrylamide (37.5:1 acrylamide/bisacrylamide) gel followed by autoradiography.

    Incubation:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. For the GC box 1 probes, 100 femtomoles of the 30-bp labeled probes containing wild-type and mutant GC box 1 (∼1 × 105 dpm) were incubated with 12 μg of uninfected or infected MeWo cell nuclear extract in a 10-μl reaction mixture in binding buffer A [40 mM HEPES (pH 7.9), 100 mM NaCl, 10 mM MgCl2 , 200 μg/ml bovine serum albumin, 12% glycerol, 0.05% NP-40, 1 mM dithiothreitol, and 3 μg poly(dI-dC)].

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Proteinase K in TE buffer (10 mM Tris HCl and 1 mM EDTA) was added to a final concentration of 100 μg per ml, and the NaCl concentration was adjusted to 0.15 M. The mixture then was incubated at 50°C for 3 h. The DNA was isolated by phenol-chloroform extraction followed by ethanol precipitation. .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Article Title: mRNA Decapping and 5′-3′ Decay Contribute to the Regulation of ABA Signaling in Arabidopsis thaliana
    Article Snippet: Northern blots were performed using random primed probes amplified from cDNA template with appropriate primers and radioactively labeled with DECAprimeTM II kit (Ambion) and [α-32 P]ATP (Hartmann Analytics), or oligoribonucleotide probes radioactively labeled with PNK (Thermo Scientific) and [γ-32 P]ATP (Hartmann Analytics). .. Two-week-old seedlings were transferred to flasks containing a buffer (1mM PIPES, pH 6.25, 1 mM sodium citrate, 1 mM KCl, 15 mM sucrose), and after a 30-min incubation, cordycepin (150 mg/L) was added.

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. One hundred femtomoles of the labeled probes containing wild-type and mutant Box D (∼1 × 105 dpm) were incubated with 15 μg of uninfected or infected MeWo cell nuclear extract or PAA-treated infected cell nuclear extract in a 10-μl reaction mixture in binding buffer [40 mM HEPES, pH 7.9, 100 mM NaCl, 10 mM MgCl2 , 200 μg/ml bovine serum albumin (BSA), 12% glycerol, 0.05% NP-40, 1 mM dithiothreitol, and 3 μg poly(dI-dC)].

    Article Title: Brome Mosaic Virus 1a Nucleoside Triphosphatase/Helicase Domain Plays Crucial Roles in Recruiting RNA Replication Templates
    Article Snippet: .. Purified proteins were incubated with 5 μCi [α-32 P]ATP in 10 μl of 50 mM Tris-HCl (pH 7.4), 5 mM MgCl2 , 5 mM DTT, 150 μM ATP, and 20 U of RNAsin (Ambion, Austin, TX) at 37°C for 30 min. .. Reactions were stopped by adding EDTA to 50 μM, and 1 μl of each reaction mixture was spotted on polyethyleneimine-cellulose thin-layer chromatography (TLC) plates (Fisher Scientific, Pittsburgh, Pa.) and developed with 0.5 M LiCl-0.5 M formic acid.

    Activity Assay:

    Article Title: Brome Mosaic Virus 1a Nucleoside Triphosphatase/Helicase Domain Plays Crucial Roles in Recruiting RNA Replication Templates
    Article Snippet: ATPase activity was assayed as previously described ( ) with minor modifications. .. Purified proteins were incubated with 5 μCi [α-32 P]ATP in 10 μl of 50 mM Tris-HCl (pH 7.4), 5 mM MgCl2 , 5 mM DTT, 150 μM ATP, and 20 U of RNAsin (Ambion, Austin, TX) at 37°C for 30 min.

    Expressing:

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: Expression of HML in strains CH2493 ( pol1-17 ) and CH2494 ( POL1 ) was examined with a PCR-generated 746-bp fragment containing the entire Yα segment of the α cassette, which recognizes both the α 1 and α 2 genes from the HML locus ( ); the PCR primers used were 5′-CTCATCTGTGATTTGTGGAT-3′ and 5′-AAGTAGTCCCATATTCCGTG-3′. .. The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion).

    Derivative Assay:

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: The blots for the experiments done with the pVO2 plasmids were probed with a 400-bp PCR product derived from pVO2 using the primers 5′-GTGCTCCTGTCGTTGAGGACCCGG-3′ and 5′-CCTCTGACTTGAGCGTCGATTTTT-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Hybridization:

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: Standard techniques were used for gel electrophoresis, RNA transfer, and hybridization of RNA samples (20 μg/lane) ( ); Quick-Hyb hybridization solution (Stratagene) was used for the hybridizations of the RNA immobilized onto Hybond-N nylon membrane (Amersham). .. The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion).

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: The DNA was digested with DpnI and EcoRI as described by Stow and Davison ( ) and analyzed by Southern blot hybridization. .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Southern Blot:

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: The DNA was digested with DpnI and EcoRI as described by Stow and Davison ( ) and analyzed by Southern blot hybridization. .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Article Title: Structure and Expression of a Dhurrinase (?-Glucosidase) from Sorghum 1
    Article Snippet: .. The purified PCR products (the 1446- and 627-bp fragments) were labeled with [α-32 P]ATP using random hexamers and the Klenow fragment of DNA polymerase (Ambion, Austin, TX) at 37°C for 1 h. The labeled fragments were used as a probe in northern- and Southern-blot analyses. .. Genomic DNA was isolated from 3- to 4-d-old seedlings as described by .

    Northern Blot:

    Article Title: A conserved mechanism of TOR-dependent RCK-mediated mRNA degradation regulates autophagy
    Article Snippet: .. DNA probes were labeled with [α-32 P]-ATP for both Southern and northern blots using the RadPrime DNA Labeling System (Invitrogen) according to the manufacturer's protocol. .. Transcript levels of indicated genes were monitored by northern blot analysis hybridized with a PCR-amplified fragment of the indicated genes using primers listed in .

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: Paragraph title: Northern blot analysis. ... The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion).

    Article Title: mRNA Decapping and 5′-3′ Decay Contribute to the Regulation of ABA Signaling in Arabidopsis thaliana
    Article Snippet: .. Northern blots were performed using random primed probes amplified from cDNA template with appropriate primers and radioactively labeled with DECAprimeTM II kit (Ambion) and [α-32 P]ATP (Hartmann Analytics), or oligoribonucleotide probes radioactively labeled with PNK (Thermo Scientific) and [γ-32 P]ATP (Hartmann Analytics). .. Membranes were hybridized overnight with radioactive probes in PerfectHyb buffer (Sigma), washed, analyzed with PhosphorImager Typhoon FLA 9000 (GE Healthcare) and quantified with ImageJ software (Molecular Dynamics). mRNA half-life measurement experiments were carried out as described (Souret et al., ).

    Article Title: Structure and Expression of a Dhurrinase (?-Glucosidase) from Sorghum 1
    Article Snippet: .. The purified PCR products (the 1446- and 627-bp fragments) were labeled with [α-32 P]ATP using random hexamers and the Klenow fragment of DNA polymerase (Ambion, Austin, TX) at 37°C for 1 h. The labeled fragments were used as a probe in northern- and Southern-blot analyses. .. Genomic DNA was isolated from 3- to 4-d-old seedlings as described by .

    Infection:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. For the GC box 1 probes, 100 femtomoles of the 30-bp labeled probes containing wild-type and mutant GC box 1 (∼1 × 105 dpm) were incubated with 12 μg of uninfected or infected MeWo cell nuclear extract in a 10-μl reaction mixture in binding buffer A [40 mM HEPES (pH 7.9), 100 mM NaCl, 10 mM MgCl2 , 200 μg/ml bovine serum albumin, 12% glycerol, 0.05% NP-40, 1 mM dithiothreitol, and 3 μg poly(dI-dC)].

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. One hundred femtomoles of the labeled probes containing wild-type and mutant Box D (∼1 × 105 dpm) were incubated with 15 μg of uninfected or infected MeWo cell nuclear extract or PAA-treated infected cell nuclear extract in a 10-μl reaction mixture in binding buffer [40 mM HEPES, pH 7.9, 100 mM NaCl, 10 mM MgCl2 , 200 μg/ml bovine serum albumin (BSA), 12% glycerol, 0.05% NP-40, 1 mM dithiothreitol, and 3 μg poly(dI-dC)].

    Generated:

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: A 571-bp fragment containing the entire ACT1 coding sequence (generated by PCR with primers 5′-CGATTTGGCCGGTAGAGATT-3′ and 5′-TAGATGGACCACTTTCGTCG-3′) was used to control for RNA loading. .. The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion).

    DNA Labeling:

    Article Title: A conserved mechanism of TOR-dependent RCK-mediated mRNA degradation regulates autophagy
    Article Snippet: .. DNA probes were labeled with [α-32 P]-ATP for both Southern and northern blots using the RadPrime DNA Labeling System (Invitrogen) according to the manufacturer's protocol. .. Transcript levels of indicated genes were monitored by northern blot analysis hybridized with a PCR-amplified fragment of the indicated genes using primers listed in .

    Polymerase Chain Reaction:

    Article Title: A conserved mechanism of TOR-dependent RCK-mediated mRNA degradation regulates autophagy
    Article Snippet: DNA probes were labeled with [α-32 P]-ATP for both Southern and northern blots using the RadPrime DNA Labeling System (Invitrogen) according to the manufacturer's protocol. .. Transcript levels of indicated genes were monitored by northern blot analysis hybridized with a PCR-amplified fragment of the indicated genes using primers listed in .

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: A 571-bp fragment containing the entire ACT1 coding sequence (generated by PCR with primers 5′-CGATTTGGCCGGTAGAGATT-3′ and 5′-TAGATGGACCACTTTCGTCG-3′) was used to control for RNA loading. .. The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion).

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. The pLitmus R62/63F fragment includes a portion (nucleotide positions 1832 to 2307) of the pLitmus R62/63F plasmid containing the pUC19 origin of replication and was designed to detect both intact DpnI-resistant linearized pLitmus R62/63F (6.9 kb) resulting from replication of the input plasmid and a 651-bp fragment resulting from the DpnI-sensitive unreplicated input plasmid.

    Article Title: Structure and Expression of a Dhurrinase (?-Glucosidase) from Sorghum 1
    Article Snippet: .. The purified PCR products (the 1446- and 627-bp fragments) were labeled with [α-32 P]ATP using random hexamers and the Klenow fragment of DNA polymerase (Ambion, Austin, TX) at 37°C for 1 h. The labeled fragments were used as a probe in northern- and Southern-blot analyses. .. Genomic DNA was isolated from 3- to 4-d-old seedlings as described by .

    Binding Assay:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: Thirty-base-pair oligonucleotide probes (IDT, Coralville, IA) containing wild-type and mutant GC box 1 elements and 40-bp probes containing wild-type and mutant YY1 and GC box 2 binding site elements were used in electrophoretic mobility shift assays (EMSAs). .. Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. One hundred femtomoles of the labeled probes containing wild-type and mutant Box D (∼1 × 105 dpm) were incubated with 15 μg of uninfected or infected MeWo cell nuclear extract or PAA-treated infected cell nuclear extract in a 10-μl reaction mixture in binding buffer [40 mM HEPES, pH 7.9, 100 mM NaCl, 10 mM MgCl2 , 200 μg/ml bovine serum albumin (BSA), 12% glycerol, 0.05% NP-40, 1 mM dithiothreitol, and 3 μg poly(dI-dC)].

    Article Title: Defining the RNA-Protein Interactions in the Trypanosome Preribosomal Complex
    Article Snippet: The 5′ ends of 5S rRNA oligonucleotides were labeled with [α-32 P]ATP (KinaseMax; Life Technologies). .. The 5S rRNA or 5S rRNA oligonucleotides and increasing concentrations of recombinant proteins were added in a total volume of 20 μl, in binding buffer (10 mM Tris [pH 7.4], 150 mM KCl, 0.1 mM EDTA, 0.1 mM DTT, 0.1% NP-40, 10% glycerol, 100 μg/ml BSA).

    Nucleic Acid Electrophoresis:

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: Standard techniques were used for gel electrophoresis, RNA transfer, and hybridization of RNA samples (20 μg/lane) ( ); Quick-Hyb hybridization solution (Stratagene) was used for the hybridizations of the RNA immobilized onto Hybond-N nylon membrane (Amersham). .. The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion).

    Mutagenesis:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: Thirty-base-pair oligonucleotide probes (IDT, Coralville, IA) containing wild-type and mutant GC box 1 elements and 40-bp probes containing wild-type and mutant YY1 and GC box 2 binding site elements were used in electrophoretic mobility shift assays (EMSAs). .. Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Fifty-one-base-pair oligonucleotide probes containing wild-type and mutant Box D elements (IDT, Coralville, IA) were used in EMSAs. .. Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Isolation:

    Article Title: A conserved mechanism of TOR-dependent RCK-mediated mRNA degradation regulates autophagy
    Article Snippet: RNA was isolated and northern blots performed as previously described . .. DNA probes were labeled with [α-32 P]-ATP for both Southern and northern blots using the RadPrime DNA Labeling System (Invitrogen) according to the manufacturer's protocol.

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: Total RNA was isolated by a phenol-freeze method ( ). .. The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion).

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Proteinase K in TE buffer (10 mM Tris HCl and 1 mM EDTA) was added to a final concentration of 100 μg per ml, and the NaCl concentration was adjusted to 0.15 M. The mixture then was incubated at 50°C for 3 h. The DNA was isolated by phenol-chloroform extraction followed by ethanol precipitation. .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Article Title: mRNA Decapping and 5′-3′ Decay Contribute to the Regulation of ABA Signaling in Arabidopsis thaliana
    Article Snippet: RNA methods Total RNA was isolated from 2-week-old seedlings using Trizol reagent (Sigma) according to the manufacturer's instructions. .. Northern blots were performed using random primed probes amplified from cDNA template with appropriate primers and radioactively labeled with DECAprimeTM II kit (Ambion) and [α-32 P]ATP (Hartmann Analytics), or oligoribonucleotide probes radioactively labeled with PNK (Thermo Scientific) and [γ-32 P]ATP (Hartmann Analytics).

    Electrophoretic Mobility Shift Assay:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: Thirty-base-pair oligonucleotide probes (IDT, Coralville, IA) containing wild-type and mutant GC box 1 elements and 40-bp probes containing wild-type and mutant YY1 and GC box 2 binding site elements were used in electrophoretic mobility shift assays (EMSAs). .. Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Purification:

    Article Title: The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase
    Article Snippet: To prepare blunt-ended double-stranded DNA (dsDNA) or dsDNA with one or two single-stranded tails, an oligonucleotide was labeled with [γ-32 P]ATP (PerklinElmer) at the 5′ end using T4 polynucleotide kinase (TaKaRa) and purified using a G-50 MicroSpin column (GE Healthcare). .. To radiolabel a 5′ biotin-labeled oligonucleotide (S14) (see Table S1), the DNA fragment was annealed to oligonucleotide S20 and filled in with [α-32 P]ATP using Klenow fragment (Thermo).

    Article Title: Structure and Expression of a Dhurrinase (?-Glucosidase) from Sorghum 1
    Article Snippet: .. The purified PCR products (the 1446- and 627-bp fragments) were labeled with [α-32 P]ATP using random hexamers and the Klenow fragment of DNA polymerase (Ambion, Austin, TX) at 37°C for 1 h. The labeled fragments were used as a probe in northern- and Southern-blot analyses. .. Genomic DNA was isolated from 3- to 4-d-old seedlings as described by .

    Article Title: Brome Mosaic Virus 1a Nucleoside Triphosphatase/Helicase Domain Plays Crucial Roles in Recruiting RNA Replication Templates
    Article Snippet: .. Purified proteins were incubated with 5 μCi [α-32 P]ATP in 10 μl of 50 mM Tris-HCl (pH 7.4), 5 mM MgCl2 , 5 mM DTT, 150 μM ATP, and 20 U of RNAsin (Ambion, Austin, TX) at 37°C for 30 min. .. Reactions were stopped by adding EDTA to 50 μM, and 1 μl of each reaction mixture was spotted on polyethyleneimine-cellulose thin-layer chromatography (TLC) plates (Fisher Scientific, Pittsburgh, Pa.) and developed with 0.5 M LiCl-0.5 M formic acid.

    Article Title: Defining the RNA-Protein Interactions in the Trypanosome Preribosomal Complex
    Article Snippet: The 5′ ends of 5S rRNA oligonucleotides were labeled with [α-32 P]ATP (KinaseMax; Life Technologies). .. Radiolabeled 5S rRNA and 5S rRNA oligonucleotides were purified using NucAway spin columns (Life Technologies).

    Article Title: Affinity Labeling of Hepatitis C Virus Replicase with a Nucleotide Analog: Identification of binding site
    Article Snippet: 37-mer double stranded RNA looped template primer and dT18 primer were labeled at 5′ using [α-32 P]ATP and T4 polynucleotide kinase (Invitrogen) according to manufacturers protocol. .. The 5′ labeled oligomers were purified by illustra ProbeQuant G-50 micro columns.

    Sequencing:

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: A 571-bp fragment containing the entire ACT1 coding sequence (generated by PCR with primers 5′-CGATTTGGCCGGTAGAGATT-3′ and 5′-TAGATGGACCACTTTCGTCG-3′) was used to control for RNA loading. .. The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion).

    Article Title: Isomorphic Emissive GTP Surrogate Facilitates Initiation and Elongation of in Vitro Transcription Reactions
    Article Snippet: Transcription Reactions with α-32 P ATP Single strand DNA templates were annealed to an 18-mer T7 RNA polymerase consensus promoter sequence in TE buffer (10 mM Tris-HCl, 1 mM EDTA, 100 mM NaCl, pH 7.8) by heating a 1:1 mixture (10 μM) at 90 °C for 3 min and cooling the solution slowly to room temperature. .. Transcription reactions were performed in 40 mM Tris-HCl buffer (pH 7.9) containing 500 nM annealed templates, 10 mM MgCl2 , 10 mM dithiothreitol (DTT), 10 mM NaCl, 2 mM spermidine, 1 U/μL RNase inhibitor (RiboLock), 1 mM GTP or 1 mM th GTP, 1 mM CTP, 1 mM UTP, 20 μM ATP, 2 μCi α-32 P ATP (800 Ci/mmol stock), and 2.5 U/μL T7 RNA polymerase (Fermentas) in a total volume of 20 μL.

    Filter-binding Assay:

    Article Title: Defining the RNA-Protein Interactions in the Trypanosome Preribosomal Complex
    Article Snippet: Paragraph title: Filter binding assay. ... The 5′ ends of 5S rRNA oligonucleotides were labeled with [α-32 P]ATP (KinaseMax; Life Technologies).

    Labeling:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: .. Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. For the GC box 1 probes, 100 femtomoles of the 30-bp labeled probes containing wild-type and mutant GC box 1 (∼1 × 105 dpm) were incubated with 12 μg of uninfected or infected MeWo cell nuclear extract in a 10-μl reaction mixture in binding buffer A [40 mM HEPES (pH 7.9), 100 mM NaCl, 10 mM MgCl2 , 200 μg/ml bovine serum albumin, 12% glycerol, 0.05% NP-40, 1 mM dithiothreitol, and 3 μg poly(dI-dC)].

    Article Title: The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase
    Article Snippet: To prepare blunt-ended double-stranded DNA (dsDNA) or dsDNA with one or two single-stranded tails, an oligonucleotide was labeled with [γ-32 P]ATP (PerklinElmer) at the 5′ end using T4 polynucleotide kinase (TaKaRa) and purified using a G-50 MicroSpin column (GE Healthcare). .. To radiolabel a 5′ biotin-labeled oligonucleotide (S14) (see Table S1), the DNA fragment was annealed to oligonucleotide S20 and filled in with [α-32 P]ATP using Klenow fragment (Thermo).

    Article Title: A conserved mechanism of TOR-dependent RCK-mediated mRNA degradation regulates autophagy
    Article Snippet: .. DNA probes were labeled with [α-32 P]-ATP for both Southern and northern blots using the RadPrime DNA Labeling System (Invitrogen) according to the manufacturer's protocol. .. Transcript levels of indicated genes were monitored by northern blot analysis hybridized with a PCR-amplified fragment of the indicated genes using primers listed in .

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: .. The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion). .. For analysis of telomeric URA3 expression, strains CH2514 ( pol1-17 ) and CH2515 ( POL1 ) were grown to early logarithmic phase at 24°C, cultures were split, and aliquots were shifted to 24 or 30°C for 2 or 5 h. The URA3 probe, which was labeled as described above, is a 911-bp Nde I- Nsi I DNA fragment of pCH1099, containing the entire coding region of the URA3 gene.

    Article Title: mRNA Decapping and 5′-3′ Decay Contribute to the Regulation of ABA Signaling in Arabidopsis thaliana
    Article Snippet: .. Northern blots were performed using random primed probes amplified from cDNA template with appropriate primers and radioactively labeled with DECAprimeTM II kit (Ambion) and [α-32 P]ATP (Hartmann Analytics), or oligoribonucleotide probes radioactively labeled with PNK (Thermo Scientific) and [γ-32 P]ATP (Hartmann Analytics). .. Membranes were hybridized overnight with radioactive probes in PerfectHyb buffer (Sigma), washed, analyzed with PhosphorImager Typhoon FLA 9000 (GE Healthcare) and quantified with ImageJ software (Molecular Dynamics). mRNA half-life measurement experiments were carried out as described (Souret et al., ).

    Article Title: Structure and Expression of a Dhurrinase (?-Glucosidase) from Sorghum 1
    Article Snippet: .. The purified PCR products (the 1446- and 627-bp fragments) were labeled with [α-32 P]ATP using random hexamers and the Klenow fragment of DNA polymerase (Ambion, Austin, TX) at 37°C for 1 h. The labeled fragments were used as a probe in northern- and Southern-blot analyses. .. Genomic DNA was isolated from 3- to 4-d-old seedlings as described by .

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: .. Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA). .. One hundred femtomoles of the labeled probes containing wild-type and mutant Box D (∼1 × 105 dpm) were incubated with 15 μg of uninfected or infected MeWo cell nuclear extract or PAA-treated infected cell nuclear extract in a 10-μl reaction mixture in binding buffer [40 mM HEPES, pH 7.9, 100 mM NaCl, 10 mM MgCl2 , 200 μg/ml bovine serum albumin (BSA), 12% glycerol, 0.05% NP-40, 1 mM dithiothreitol, and 3 μg poly(dI-dC)].

    Article Title: Defining the RNA-Protein Interactions in the Trypanosome Preribosomal Complex
    Article Snippet: .. The 5′ ends of 5S rRNA oligonucleotides were labeled with [α-32 P]ATP (KinaseMax; Life Technologies). .. Radiolabeled 5S rRNA and 5S rRNA oligonucleotides were purified using NucAway spin columns (Life Technologies).

    Article Title: Affinity Labeling of Hepatitis C Virus Replicase with a Nucleotide Analog: Identification of binding site
    Article Snippet: .. 37-mer double stranded RNA looped template primer and dT18 primer were labeled at 5′ using [α-32 P]ATP and T4 polynucleotide kinase (Invitrogen) according to manufacturers protocol. .. The 5′ labeled oligomers were purified by illustra ProbeQuant G-50 micro columns.

    IA:

    Article Title: Cellular Transcription Factors Sp1 and Sp3 Suppress Varicella-Zoster Virus Origin-Dependent DNA Replication ▿
    Article Snippet: Thirty-base-pair oligonucleotide probes (IDT, Coralville, IA) containing wild-type and mutant GC box 1 elements and 40-bp probes containing wild-type and mutant YY1 and GC box 2 binding site elements were used in electrophoretic mobility shift assays (EMSAs). .. Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Fifty-one-base-pair oligonucleotide probes containing wild-type and mutant Box D elements (IDT, Coralville, IA) were used in EMSAs. .. Probes were end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Random Primed:

    Article Title: mRNA Decapping and 5′-3′ Decay Contribute to the Regulation of ABA Signaling in Arabidopsis thaliana
    Article Snippet: .. Northern blots were performed using random primed probes amplified from cDNA template with appropriate primers and radioactively labeled with DECAprimeTM II kit (Ambion) and [α-32 P]ATP (Hartmann Analytics), or oligoribonucleotide probes radioactively labeled with PNK (Thermo Scientific) and [γ-32 P]ATP (Hartmann Analytics). .. Membranes were hybridized overnight with radioactive probes in PerfectHyb buffer (Sigma), washed, analyzed with PhosphorImager Typhoon FLA 9000 (GE Healthcare) and quantified with ImageJ software (Molecular Dynamics). mRNA half-life measurement experiments were carried out as described (Souret et al., ).

    ATPase Assay:

    Article Title: Brome Mosaic Virus 1a Nucleoside Triphosphatase/Helicase Domain Plays Crucial Roles in Recruiting RNA Replication Templates
    Article Snippet: Paragraph title: ATPase assay. ... Purified proteins were incubated with 5 μCi [α-32 P]ATP in 10 μl of 50 mM Tris-HCl (pH 7.4), 5 mM MgCl2 , 5 mM DTT, 150 μM ATP, and 20 U of RNAsin (Ambion, Austin, TX) at 37°C for 30 min.

    Plasmid Preparation:

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: The pVO2 fragment includes a portion (nucleotide positions 1465 to 1864) of the pVO2 plasmid containing the Col E1 origin of replication and was designed to detect both intact DpnI-resistant linearized pVO2 (3.9 kb) resulting from the replication of the input plasmid and an 872-bp fragment resulting from the DpnI-sensitive unreplicated input plasmid. .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Software:

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion). .. Quantitation was performed with a Molecular Dynamics STORM PhosphorImager and ImageQuaNT software.

    Article Title: mRNA Decapping and 5′-3′ Decay Contribute to the Regulation of ABA Signaling in Arabidopsis thaliana
    Article Snippet: Northern blots were performed using random primed probes amplified from cDNA template with appropriate primers and radioactively labeled with DECAprimeTM II kit (Ambion) and [α-32 P]ATP (Hartmann Analytics), or oligoribonucleotide probes radioactively labeled with PNK (Thermo Scientific) and [γ-32 P]ATP (Hartmann Analytics). .. Membranes were hybridized overnight with radioactive probes in PerfectHyb buffer (Sigma), washed, analyzed with PhosphorImager Typhoon FLA 9000 (GE Healthcare) and quantified with ImageJ software (Molecular Dynamics). mRNA half-life measurement experiments were carried out as described (Souret et al., ).

    Recombinant:

    Article Title: Defining the RNA-Protein Interactions in the Trypanosome Preribosomal Complex
    Article Snippet: The 5′ ends of 5S rRNA oligonucleotides were labeled with [α-32 P]ATP (KinaseMax; Life Technologies). .. The 5S rRNA or 5S rRNA oligonucleotides and increasing concentrations of recombinant proteins were added in a total volume of 20 μl, in binding buffer (10 mM Tris [pH 7.4], 150 mM KCl, 0.1 mM EDTA, 0.1 mM DTT, 0.1% NP-40, 10% glycerol, 100 μg/ml BSA).

    Agarose Gel Electrophoresis:

    Article Title: Structure and Expression of a Dhurrinase (?-Glucosidase) from Sorghum 1
    Article Snippet: After PCR, the amplification products were electrophoresed through a low-melting agarose gel (1%), excised, and purified using the Magic PCR Prep system according to the procedure provided by the manufacturer (Promega). .. The purified PCR products (the 1446- and 627-bp fragments) were labeled with [α-32 P]ATP using random hexamers and the Klenow fragment of DNA polymerase (Ambion, Austin, TX) at 37°C for 1 h. The labeled fragments were used as a probe in northern- and Southern-blot analyses.

    In Vitro:

    Article Title: Defining the RNA-Protein Interactions in the Trypanosome Preribosomal Complex
    Article Snippet: T. brucei 5S rRNA was transcribed in vitro in the presence of [α-32 P]UTP as described previously ( , ). .. The 5′ ends of 5S rRNA oligonucleotides were labeled with [α-32 P]ATP (KinaseMax; Life Technologies).

    Ethanol Precipitation:

    Article Title: The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase
    Article Snippet: To radiolabel a 5′ biotin-labeled oligonucleotide (S14) (see Table S1), the DNA fragment was annealed to oligonucleotide S20 and filled in with [α-32 P]ATP using Klenow fragment (Thermo). .. Following ethanol precipitation, the DNA was dissolved in Tris-EDTA (TE) buffer.

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Proteinase K in TE buffer (10 mM Tris HCl and 1 mM EDTA) was added to a final concentration of 100 μg per ml, and the NaCl concentration was adjusted to 0.15 M. The mixture then was incubated at 50°C for 3 h. The DNA was isolated by phenol-chloroform extraction followed by ethanol precipitation. .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Quantitation Assay:

    Article Title: The Function of DNA Polymerase ? at Telomeric G Tails Is Important for Telomere Homeostasis
    Article Snippet: The probes were labeled with [α-32 P]ATP by random priming (DECAprime; Ambion). .. Quantitation was performed with a Molecular Dynamics STORM PhosphorImager and ImageQuaNT software.

    Concentration Assay:

    Article Title: A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein ▿
    Article Snippet: Proteinase K in TE buffer (10 mM Tris HCl and 1 mM EDTA) was added to a final concentration of 100 μg per ml, and the NaCl concentration was adjusted to 0.15 M. The mixture then was incubated at 50°C for 3 h. The DNA was isolated by phenol-chloroform extraction followed by ethanol precipitation. .. The blots for the experiments done with the pLitmus R62/63F plasmids were probed with a 476-bp PCR product prepared from pLitmus R62/63F using the primers 5′-TAGGCCACCACTTCAAGAACTCTGT-3′ and 5′-AGCAAAAGGCCAGCAAAAGGCCAGG-3′ and end labeled with [α-32 P]ATP using T4 kinase (Invitrogen, Carlsbad, CA).

    Article Title: Defining the RNA-Protein Interactions in the Trypanosome Preribosomal Complex
    Article Snippet: The 5′ ends of 5S rRNA oligonucleotides were labeled with [α-32 P]ATP (KinaseMax; Life Technologies). .. A constant concentration of 5S rRNA (equivalent to 10,000 dpm and always lower than 0.5 nM) was used for all reactions.

    Thin Layer Chromatography:

    Article Title: Brome Mosaic Virus 1a Nucleoside Triphosphatase/Helicase Domain Plays Crucial Roles in Recruiting RNA Replication Templates
    Article Snippet: Purified proteins were incubated with 5 μCi [α-32 P]ATP in 10 μl of 50 mM Tris-HCl (pH 7.4), 5 mM MgCl2 , 5 mM DTT, 150 μM ATP, and 20 U of RNAsin (Ambion, Austin, TX) at 37°C for 30 min. .. Reactions were stopped by adding EDTA to 50 μM, and 1 μl of each reaction mixture was spotted on polyethyleneimine-cellulose thin-layer chromatography (TLC) plates (Fisher Scientific, Pittsburgh, Pa.) and developed with 0.5 M LiCl-0.5 M formic acid.

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    Thermo Fisher α 32 p atp
    α 32 P Atp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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