polynucleotide kinase buffer  (New England Biolabs)


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    Name:
    T4 Polynucleotide Kinase Reaction Buffer
    Description:
    T4 Polynucleotide Kinase Reaction Buffer 4 0 ml
    Catalog Number:
    b0201s
    Price:
    24
    Size:
    4 0 ml
    Category:
    Buffers
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    Structured Review

    New England Biolabs polynucleotide kinase buffer
    T4 Polynucleotide Kinase Reaction Buffer
    T4 Polynucleotide Kinase Reaction Buffer 4 0 ml
    https://www.bioz.com/result/polynucleotide kinase buffer/product/New England Biolabs
    Average 99 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    polynucleotide kinase buffer - by Bioz Stars, 2020-07
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3
    Article Snippet: .. CRISPR/Cas9-mediated deletion of PINCR gRNAs targeting the 5’ and 3’ ends of PINCR were designed using Zifit software ( http://zifit.partners.org/ZiFiT/ ) and were cloned into U6-gRNA vector ( ) having BsmB1 restriction enzyme site. gRNAs sequence information is provided in . gRNA oligos were ligated and phosphorylated using T4 ligation buffer (NEB) and T4 Polynucleotide Kinase (NEB) using a thermocycler with following parameter: 37°C for 30 min, 95°C for 5 min and then ramp down to 25°C at 5 °C/min. .. Annealed oligos were ligated with BsmB1 digested U6-gRNA vector (2.9 kb fragment) using quick DNA ligase (NEB).

    Radioactivity:

    Article Title: Electrical Probes of DNA-Binding Proteins
    Article Snippet: .. Solvents and Reagents : T4 polynucleotide kinase (New England Biolabs) T4 buffer (New England Biolabs) 10pmol ssDNA with free 5’ ends γ−32 P ATP (Perkin Elmer; 3000–6000 Ci/mmol) MQ water Ethylenediaminetetraacetic acid (EDTA) Instruments and Supplies : Benchtop incubators MicroBioSpin6 columns (BioRad) Tabletop centrifuge 1.5mL Eppendorf tubes Procedure : Prepare reactions mixes (50μL) in Eppendorf tubes by adding DNA, 5μL 10× concentrated T4 buffer, and MQ water; keep on ice Thaw 32 P-labeled ATP, and add 40μCi to each reaction tube (All steps involving radioactivity should be carried out behind a Lucite shield!) .. Add 1.0μL T4 polynucleotide kinase (5units) to each tube, and start reactions by sealing the tube and incubating at 37°C for 30min Stop reactions by adding EDTA to a final concentration of 10m M , and heat inactivate the kinase by incubation at 85°C for 10min Isolate DNA by adding quenched reactions to a MicroBioSpin6 column and spinning for 4min at 1000 × g

    Ligation:

    Article Title: Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3
    Article Snippet: .. CRISPR/Cas9-mediated deletion of PINCR gRNAs targeting the 5’ and 3’ ends of PINCR were designed using Zifit software ( http://zifit.partners.org/ZiFiT/ ) and were cloned into U6-gRNA vector ( ) having BsmB1 restriction enzyme site. gRNAs sequence information is provided in . gRNA oligos were ligated and phosphorylated using T4 ligation buffer (NEB) and T4 Polynucleotide Kinase (NEB) using a thermocycler with following parameter: 37°C for 30 min, 95°C for 5 min and then ramp down to 25°C at 5 °C/min. .. Annealed oligos were ligated with BsmB1 digested U6-gRNA vector (2.9 kb fragment) using quick DNA ligase (NEB).

    Article Title: TAZ induces lung cancer stem cell properties and tumorigenesis by up-regulating ALDH1A1
    Article Snippet: .. Oligos were phosphorylated and annealed by T4 polynucleotide kinase (PNK, NEB, #M0201S) in T4 ligation Buffer (NEB) following a thermocycler running at 37°C for 30 minutes, 90°C for 5 minutes and then ramp down to 25°C at 5°C per minute. .. Ligation was catalyzed by mixing annealed oligos and digested LentiCRIPSR v1 vector with Quick Ligase in Quick Ligase Buffer (NEB, #M2200S), followed by transformation into Stbl3 bacteria.

    Article Title: Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis
    Article Snippet: .. SgRNA-specifying oligos were phosphorylated and annealed using the following conditions: sgRNA sequence oligo (100 μM); sgRNA reverse complement oligo (100 μM); T4 ligation buffer (1X); and T4 polynucleotide kinase (5 units) with the following temperature conditions (New England Biolabs): 37°C for 30 minutes; 95°C for 5 minutes; and then ramp down to 25°C at 5°C min−1 . .. Annealed oligos were ligated into lentiGuide in a 1:3 ratio (vector: insert) using T4 ligation buffer (1X) and T4 DNA ligase (750 units).

    Article Title: A microfluidic oligonucleotide synthesizer
    Article Snippet: .. Ligation assembly of DNA constructs Each oligonucleotide strand (8 µl, 5 µM) was mixed with 1 µl of T4 polynucleotide kinase reaction buffer (New England Biolabs) and 1 µl of 10 mM ATP, then cooled to 4°C. ..

    Construct:

    Article Title: A microfluidic oligonucleotide synthesizer
    Article Snippet: .. Ligation assembly of DNA constructs Each oligonucleotide strand (8 µl, 5 µM) was mixed with 1 µl of T4 polynucleotide kinase reaction buffer (New England Biolabs) and 1 µl of 10 mM ATP, then cooled to 4°C. ..

    Incubation:

    Article Title: A Novel AT-Rich DNA Recognition Mechanism for Bacterial Xenogeneic Silencer MvaT
    Article Snippet: .. 400 nM DNA fragment, 4 μL γ-32 P ATP (3,000 Ci/mmol, 10 mCi/mL, Perkin Elmer), 1x polynucleotide kinase buffer and T4 polynucleotide kinase enzyme (New England Biolabs) were incubated in a total of 40 μL at 37°C for 30 min. .. The reaction was stopped by the addition of 1 μL of 0.5 M EDTA and excess radioisotopes were removed using a G-25 spin column (GE Healthcare Life Sciences).

    CRISPR:

    Article Title: Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3
    Article Snippet: .. CRISPR/Cas9-mediated deletion of PINCR gRNAs targeting the 5’ and 3’ ends of PINCR were designed using Zifit software ( http://zifit.partners.org/ZiFiT/ ) and were cloned into U6-gRNA vector ( ) having BsmB1 restriction enzyme site. gRNAs sequence information is provided in . gRNA oligos were ligated and phosphorylated using T4 ligation buffer (NEB) and T4 Polynucleotide Kinase (NEB) using a thermocycler with following parameter: 37°C for 30 min, 95°C for 5 min and then ramp down to 25°C at 5 °C/min. .. Annealed oligos were ligated with BsmB1 digested U6-gRNA vector (2.9 kb fragment) using quick DNA ligase (NEB).

    Sequencing:

    Article Title: Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3
    Article Snippet: .. CRISPR/Cas9-mediated deletion of PINCR gRNAs targeting the 5’ and 3’ ends of PINCR were designed using Zifit software ( http://zifit.partners.org/ZiFiT/ ) and were cloned into U6-gRNA vector ( ) having BsmB1 restriction enzyme site. gRNAs sequence information is provided in . gRNA oligos were ligated and phosphorylated using T4 ligation buffer (NEB) and T4 Polynucleotide Kinase (NEB) using a thermocycler with following parameter: 37°C for 30 min, 95°C for 5 min and then ramp down to 25°C at 5 °C/min. .. Annealed oligos were ligated with BsmB1 digested U6-gRNA vector (2.9 kb fragment) using quick DNA ligase (NEB).

    Article Title: Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis
    Article Snippet: .. SgRNA-specifying oligos were phosphorylated and annealed using the following conditions: sgRNA sequence oligo (100 μM); sgRNA reverse complement oligo (100 μM); T4 ligation buffer (1X); and T4 polynucleotide kinase (5 units) with the following temperature conditions (New England Biolabs): 37°C for 30 minutes; 95°C for 5 minutes; and then ramp down to 25°C at 5°C min−1 . .. Annealed oligos were ligated into lentiGuide in a 1:3 ratio (vector: insert) using T4 ligation buffer (1X) and T4 DNA ligase (750 units).

    Plasmid Preparation:

    Article Title: Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3
    Article Snippet: .. CRISPR/Cas9-mediated deletion of PINCR gRNAs targeting the 5’ and 3’ ends of PINCR were designed using Zifit software ( http://zifit.partners.org/ZiFiT/ ) and were cloned into U6-gRNA vector ( ) having BsmB1 restriction enzyme site. gRNAs sequence information is provided in . gRNA oligos were ligated and phosphorylated using T4 ligation buffer (NEB) and T4 Polynucleotide Kinase (NEB) using a thermocycler with following parameter: 37°C for 30 min, 95°C for 5 min and then ramp down to 25°C at 5 °C/min. .. Annealed oligos were ligated with BsmB1 digested U6-gRNA vector (2.9 kb fragment) using quick DNA ligase (NEB).

    Software:

    Article Title: Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3
    Article Snippet: .. CRISPR/Cas9-mediated deletion of PINCR gRNAs targeting the 5’ and 3’ ends of PINCR were designed using Zifit software ( http://zifit.partners.org/ZiFiT/ ) and were cloned into U6-gRNA vector ( ) having BsmB1 restriction enzyme site. gRNAs sequence information is provided in . gRNA oligos were ligated and phosphorylated using T4 ligation buffer (NEB) and T4 Polynucleotide Kinase (NEB) using a thermocycler with following parameter: 37°C for 30 min, 95°C for 5 min and then ramp down to 25°C at 5 °C/min. .. Annealed oligos were ligated with BsmB1 digested U6-gRNA vector (2.9 kb fragment) using quick DNA ligase (NEB).

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  • 99
    New England Biolabs t4 pnk buffer
    Manipulation of 5′ppp-Triggered Interferon Response HEK293 cells were transfected with poly(I:C) or several tsli-siRNAs. The final concentration of 10 nM for each RNAi reagent was used in transfection for qPCR assay. Gene expression level changes in OAS1, IRF9, CDKL, and IFNB relative to GAPDH were measured by qPCR. (A) Mild interferon response was observed from all four tsli-siRNAs, with tsli-RRM2 having the strongest response among them. G-tsli-Stat3 exhibited a much stronger response than tsli-Stat3, and GG-tsli-Stat3 reversed this effect to some extent. (B) CIP treatment minimized the strong interferon response by G-tsli-Stat3. (C) CIP treatment minimized and <t>T4</t> PNK treatment elevated the interferon response by tsli-RRM2. Fold changes in gene expression were normalized to untreated HEK293 cells. Details of qPCR procedure and results calculation were provided in the Materials and Methods . Error bars indicate SD.
    T4 Pnk Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 pnk buffer/product/New England Biolabs
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    t4 pnk buffer - by Bioz Stars, 2020-07
    99/100 stars
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    Manipulation of 5′ppp-Triggered Interferon Response HEK293 cells were transfected with poly(I:C) or several tsli-siRNAs. The final concentration of 10 nM for each RNAi reagent was used in transfection for qPCR assay. Gene expression level changes in OAS1, IRF9, CDKL, and IFNB relative to GAPDH were measured by qPCR. (A) Mild interferon response was observed from all four tsli-siRNAs, with tsli-RRM2 having the strongest response among them. G-tsli-Stat3 exhibited a much stronger response than tsli-Stat3, and GG-tsli-Stat3 reversed this effect to some extent. (B) CIP treatment minimized the strong interferon response by G-tsli-Stat3. (C) CIP treatment minimized and T4 PNK treatment elevated the interferon response by tsli-RRM2. Fold changes in gene expression were normalized to untreated HEK293 cells. Details of qPCR procedure and results calculation were provided in the Materials and Methods . Error bars indicate SD.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: A Simple and Cost-Effective Approach for In Vitro Production of Sliced siRNAs as Potent Triggers for RNAi

    doi: 10.1016/j.omtn.2017.07.008

    Figure Lengend Snippet: Manipulation of 5′ppp-Triggered Interferon Response HEK293 cells were transfected with poly(I:C) or several tsli-siRNAs. The final concentration of 10 nM for each RNAi reagent was used in transfection for qPCR assay. Gene expression level changes in OAS1, IRF9, CDKL, and IFNB relative to GAPDH were measured by qPCR. (A) Mild interferon response was observed from all four tsli-siRNAs, with tsli-RRM2 having the strongest response among them. G-tsli-Stat3 exhibited a much stronger response than tsli-Stat3, and GG-tsli-Stat3 reversed this effect to some extent. (B) CIP treatment minimized the strong interferon response by G-tsli-Stat3. (C) CIP treatment minimized and T4 PNK treatment elevated the interferon response by tsli-RRM2. Fold changes in gene expression were normalized to untreated HEK293 cells. Details of qPCR procedure and results calculation were provided in the Materials and Methods . Error bars indicate SD.

    Article Snippet: The protocol was modified as follows when CIP or T4 PNK treatment is necessary: (1) for CIP treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of CIP, 4 μL of 10× CutSmart buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min; and (2) for T4 PNK treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of T4 PNK, 4 μL of 10× T4 PNK buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min. All T7 in vitro transcription products were purified by Micro Bio-Spin P-30 Gel Columns, Tris Buffer, from Bio-Rad.

    Techniques: Transfection, Concentration Assay, Real-time Polymerase Chain Reaction, Expressing

    Read distribution of ex‑mRNA reads across the full-length mRNA transcripts. ( A and B ) Read coverage for the hemoglobin A2 transcript ( A ) and the albumin transcript ( B ) by sample type for untreated and T4 PNK end-treated samples. Exon boundaries (HBA2: 3 exons, ALB: 15 exons) are indicated by alternating intensities of gray, and UTRs are distinguished from CDS by thinner bars. ( C ) Metagene analysis with relative read coverage (percentage) across 5′ UTRs, CDSs, and 3′ UTRs for untreated and PNK-treated samples as well as corresponding data obtained after 100 random simulations (across an average of 2342–3500 captured transcripts for untreated samples and an average of 12,789–16,487 captured transcripts for PNK-treated samples, depending on sample type). Shown are results from n = 6 individual samples per condition.

    Journal: JCI Insight

    Article Title: Detection of circulating extracellular mRNAs by modified small-RNA-sequencing analysis

    doi: 10.1172/jci.insight.127317

    Figure Lengend Snippet: Read distribution of ex‑mRNA reads across the full-length mRNA transcripts. ( A and B ) Read coverage for the hemoglobin A2 transcript ( A ) and the albumin transcript ( B ) by sample type for untreated and T4 PNK end-treated samples. Exon boundaries (HBA2: 3 exons, ALB: 15 exons) are indicated by alternating intensities of gray, and UTRs are distinguished from CDS by thinner bars. ( C ) Metagene analysis with relative read coverage (percentage) across 5′ UTRs, CDSs, and 3′ UTRs for untreated and PNK-treated samples as well as corresponding data obtained after 100 random simulations (across an average of 2342–3500 captured transcripts for untreated samples and an average of 12,789–16,487 captured transcripts for PNK-treated samples, depending on sample type). Shown are results from n = 6 individual samples per condition.

    Article Snippet: To half of the eluted exRNA, i.e., 14 μl, we added 6 μl of a master mix corresponding to the equivalent of 2 μl ×10 T4 PNK buffer, 2 μl 10 mM ATP, 1 μl RNase-free water, and 1 μl T4 PNK (NEB, catalog M0201S) for a final reaction volume of 20 μl in a 1.5 ml siliconized microcentrifuge tube.

    Techniques:

    Treatment of total extracellular RNA with T4 polynucleotide kinase followed by small-RNA-sequencing. ( A ) Total RNA was isolated from 450 μl serum or platelet-depleted EDTA, acid citrate dextrose (ACD), and heparin plasma from 6 healthy individuals and purified using silica-based spin columns. Half of the RNA was treated with T4 polynucleotide kinase (T4 PNK) and repurified (PNK treated), and multiplexed small-RNA-sequencing (sRNA-seq) libraries were prepared separately for the untreated (libraries 1 and 3) and PNK-treated RNA (libraries 2 and 4). ( B ) Differences in read annotation in the 4 sample types for untreated RNA and PNK-treated RNA using initial annotation settings (reads 12–42 nt, up to 2 mismatches, multimapping). ( C ) Differences in ex‑mRNA capture between untreated and PNK-treated RNA using final annotation criteria (reads  > 15 nt, no mismatch and up to 2 mapping locations). Box plots show the median and first and third quartiles (bottom and top hinges). Whiskers extend at most ×1.5 interquartile range from the hinges; any data outside this are shown as individual outlier points. Shown are results from  n  = 6 individual samples per condition.

    Journal: JCI Insight

    Article Title: Detection of circulating extracellular mRNAs by modified small-RNA-sequencing analysis

    doi: 10.1172/jci.insight.127317

    Figure Lengend Snippet: Treatment of total extracellular RNA with T4 polynucleotide kinase followed by small-RNA-sequencing. ( A ) Total RNA was isolated from 450 μl serum or platelet-depleted EDTA, acid citrate dextrose (ACD), and heparin plasma from 6 healthy individuals and purified using silica-based spin columns. Half of the RNA was treated with T4 polynucleotide kinase (T4 PNK) and repurified (PNK treated), and multiplexed small-RNA-sequencing (sRNA-seq) libraries were prepared separately for the untreated (libraries 1 and 3) and PNK-treated RNA (libraries 2 and 4). ( B ) Differences in read annotation in the 4 sample types for untreated RNA and PNK-treated RNA using initial annotation settings (reads 12–42 nt, up to 2 mismatches, multimapping). ( C ) Differences in ex‑mRNA capture between untreated and PNK-treated RNA using final annotation criteria (reads > 15 nt, no mismatch and up to 2 mapping locations). Box plots show the median and first and third quartiles (bottom and top hinges). Whiskers extend at most ×1.5 interquartile range from the hinges; any data outside this are shown as individual outlier points. Shown are results from n = 6 individual samples per condition.

    Article Snippet: To half of the eluted exRNA, i.e., 14 μl, we added 6 μl of a master mix corresponding to the equivalent of 2 μl ×10 T4 PNK buffer, 2 μl 10 mM ATP, 1 μl RNase-free water, and 1 μl T4 PNK (NEB, catalog M0201S) for a final reaction volume of 20 μl in a 1.5 ml siliconized microcentrifuge tube.

    Techniques: RNA Sequencing Assay, Isolation, Purification

    Endonucleolytically cleaved 5’-OH RNAs are phosphorylated by Trl1. a 8% PAGE followed by northern blot analysis using probe prA. Levels of 3’-NGD RNA fragments in trl1/dom34 cells compared with those from TRL1/dom34 cells. b B1 and B4 RNA quantification relative to 5S rRNA from three independent experiments as shown in ( a ). c 12% PAGE followed by northern blot analysis using probe prA. Treatment using T4 PNK to determine 5’-OH and 5’-P B4 RNA positions in the indicated strains. One-fourth of trl1/dom34 total RNA treated was loaded to limit scan saturation and allow TRL1/dom34 B4 RNA detection. The 5S rRNA served as a loading control. d As in Fig. 3a , Xrn1 digestion of total RNA extracts from trl1/dom34 mutant cells in the presence or absence of T4 PNK treatment in vitro . A minor band detected in trl1 is indicated by an asterisk (see also Supplementary Fig. 5 in which this band is detectable in TRL1 cells). Error bars indicate standard deviation (s.d) calculated from three independent experiments. Source data are provided as a Source Data file.

    Journal: bioRxiv

    Article Title: No-Go Decay mRNA cleavage in the ribosome exit tunnel produces 5’-OH ends phosphorylated by Trl1

    doi: 10.1101/465633

    Figure Lengend Snippet: Endonucleolytically cleaved 5’-OH RNAs are phosphorylated by Trl1. a 8% PAGE followed by northern blot analysis using probe prA. Levels of 3’-NGD RNA fragments in trl1/dom34 cells compared with those from TRL1/dom34 cells. b B1 and B4 RNA quantification relative to 5S rRNA from three independent experiments as shown in ( a ). c 12% PAGE followed by northern blot analysis using probe prA. Treatment using T4 PNK to determine 5’-OH and 5’-P B4 RNA positions in the indicated strains. One-fourth of trl1/dom34 total RNA treated was loaded to limit scan saturation and allow TRL1/dom34 B4 RNA detection. The 5S rRNA served as a loading control. d As in Fig. 3a , Xrn1 digestion of total RNA extracts from trl1/dom34 mutant cells in the presence or absence of T4 PNK treatment in vitro . A minor band detected in trl1 is indicated by an asterisk (see also Supplementary Fig. 5 in which this band is detectable in TRL1 cells). Error bars indicate standard deviation (s.d) calculated from three independent experiments. Source data are provided as a Source Data file.

    Article Snippet: NEB Buffer 3 was replaced by T4 PNK buffer (NEB) in kinase assays in the presence or absence of Xrn1 (Figs and ).

    Techniques: Polyacrylamide Gel Electrophoresis, Northern Blot, RNA Detection, Mutagenesis, In Vitro, Standard Deviation

    Endonucleolytically cleaved 5′-OH RNAs are phosphorylated by Trl1. a 8% PAGE followed by northern blot analysis using probe prA. Levels of 3′-NGD RNA fragments in trl1/dom34 cells compared with those from TRL1/dom34 cells. b B1 and B4 RNA quantification relative to 5S rRNA from three independent experiments as shown in a . c 12% PAGE followed by northern blot analysis using probe prA. Treatment using T4 PNK to determine 5’-OH and 5’-P B4 RNA positions in the indicated strains. One-fourth of trl1/dom34 total RNA treated was loaded to limit scan saturation and allow TRL1/dom34 B4 RNA detection. The 5S rRNA served as a loading control. d As in Fig. 3a , Xrn1 digestion of total RNA extracts from trl1/dom34 mutant cells in the presence or absence of T4 PNK treatment in vitro. A minor band detected in trl1 is indicated by an asterisk (see also Supplementary Fig. 5 in which this band is detectable in TRL1 cells). Error bars indicate standard deviation (s.d.) calculated from three independent experiments. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: No-Go Decay mRNA cleavage in the ribosome exit tunnel produces 5′-OH ends phosphorylated by Trl1

    doi: 10.1038/s41467-019-13991-9

    Figure Lengend Snippet: Endonucleolytically cleaved 5′-OH RNAs are phosphorylated by Trl1. a 8% PAGE followed by northern blot analysis using probe prA. Levels of 3′-NGD RNA fragments in trl1/dom34 cells compared with those from TRL1/dom34 cells. b B1 and B4 RNA quantification relative to 5S rRNA from three independent experiments as shown in a . c 12% PAGE followed by northern blot analysis using probe prA. Treatment using T4 PNK to determine 5’-OH and 5’-P B4 RNA positions in the indicated strains. One-fourth of trl1/dom34 total RNA treated was loaded to limit scan saturation and allow TRL1/dom34 B4 RNA detection. The 5S rRNA served as a loading control. d As in Fig. 3a , Xrn1 digestion of total RNA extracts from trl1/dom34 mutant cells in the presence or absence of T4 PNK treatment in vitro. A minor band detected in trl1 is indicated by an asterisk (see also Supplementary Fig. 5 in which this band is detectable in TRL1 cells). Error bars indicate standard deviation (s.d.) calculated from three independent experiments. Source data are provided as a Source Data file.

    Article Snippet: NEB Buffer 3 was replaced by T4 PNK buffer (NEB) in kinase assays in the presence or absence of Xrn1 (Figs. a and ).

    Techniques: Polyacrylamide Gel Electrophoresis, Northern Blot, RNA Detection, Mutagenesis, In Vitro, Standard Deviation