human placenta rnase inhibitor  (New England Biolabs)


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    Name:
    RNase Inhibitor Human Placenta
    Description:
    RNase Inhibitor Human Placenta 10 000 units
    Catalog Number:
    M0307L
    Price:
    317
    Size:
    10 000 units
    Category:
    Enzyme Inhibitors
    Score:
    85
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    Structured Review

    New England Biolabs human placenta rnase inhibitor
    RNase Inhibitor Human Placenta
    RNase Inhibitor Human Placenta 10 000 units
    https://www.bioz.com/result/human placenta rnase inhibitor/product/New England Biolabs
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human placenta rnase inhibitor - by Bioz Stars, 2019-10
    98/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Following centrifugation for 10 min at 13,000 rpm in an Eppendorf 5424 R centrifuge (Eppendorf; Cat. No. 5404000138) at 4 °C, the precipitated DNA was washed with 70% ice-cold ethanol before air drying and re-suspending in RNAse-free water. .. Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S).

    Amplification:

    Article Title: Radical SAM-Mediated Methylation of Ribosomal RNA
    Article Snippet: In vitro transcription reactions of 23S rRNA fragments are carried out in the following manner: in the first step, a desired fragment of the DNA template is amplified using a plasmid pKK3535 that encodes a 23S rRNA gene ( ). .. In a typical in vitro transcription reaction, 20 μg of DNA template is dissolved in 500 μl solution containing T7 buffer (40 m M Tris–HCl, pH 7.9, 16 m M MgCl2 , 2 mM spermidine), 20 m M DTT, 3.2 m M of each of rNTP, 0.2 U/μl RNase inhibitor from human placenta, and 3 U/μl T7 RNA polymerase (New England BioLabs).

    Article Title: Barriers to transmission of transcriptional noise in a c-fos c-jun pathway
    Article Snippet: To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents. .. To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents.

    Synthesized:

    Article Title: The Evolutionarily Conserved Cassette Exon 7b Drives ERG's Oncogenic Properties
    Article Snippet: Total RNA was extracted using the total RNA isolation mini kit (Agilent Technologies Ltd.). .. All samples were treated with DNAse on the columns using RNase-free DNase I provided in the kit. cDNA was synthesized from 0.2-1 μg of total RNA using 200 U MuLV reverse transcriptase (New England Biolabs), 40 U RNase inhibitor (human placenta) (New England Biolabs), 0.5 mM dNTP, 25 μM oligo-dT primers, and 10× reverse transcriptase buffer (500 mM Tris–HCl pH 8.3, 750 mM KCl, 30 mM MgCl2 , 100 mM DTT) (New England Biolabs) in a final reaction volume 20 μl with added nuclease-free water as required (Qiagen). .. Hot Start Taq 2× master mix (New England Biolabs) was used for standard PCR.

    Article Title: RNA Sequences Required for the Noncoding Function of oskar RNA also Mediate Regulation of Oskar Protein Expression by Bicoid Stability Factor
    Article Snippet: The samples were incubated for 1 hr at 37 °C with 2.5 U of RNase H (NEB) and 20 U of RNase Inhibitor, Human Placenta (HPRI, NEB) in 50 μl of RNase H buffer (NEB). .. For circularization, decapped RNA (1.5 μg) was incubated with 10 U of T4 RNA ligase (NEB) and 20 U of HPRI in 200 μl T4 RNA ligase buffer, 0.1 mM ATP, and 10% PEG8000 for over 16 hr at 16 °C.

    Quantitative RT-PCR:

    Article Title: Stabilization of Urinary MicroRNAs by Association with Exosomes and Argonaute 2 Protein
    Article Snippet: Paragraph title: 5.3. Reverse Transcription-Quantitative PCR (RT-qPCR) Analysis ... The RT master mix for one reaction was composed of: 4.25 μL of water, 1.5 μL of 10 × RT Buffer, 0.15 μL of 100 mM dNTP, 0.1 μL of 40 U/μL RNase Inhibitor (M0307S, New England BioLabs® Inc., Ipswich, MA, USA), 1 μL of 50 U/μL MultiScribe Reverse Transcriptase and 3 μL of 5 × RT-primer specific for each miRNA.

    Real-time Polymerase Chain Reaction:

    Article Title: RNA Sequences Required for the Noncoding Function of oskar RNA also Mediate Regulation of Oskar Protein Expression by Bicoid Stability Factor
    Article Snippet: RNAs were prepared as for qPCR. .. The samples were incubated for 1 hr at 37 °C with 2.5 U of RNase H (NEB) and 20 U of RNase Inhibitor, Human Placenta (HPRI, NEB) in 50 μl of RNase H buffer (NEB).

    Article Title: Stabilization of Urinary MicroRNAs by Association with Exosomes and Argonaute 2 Protein
    Article Snippet: The RT master mix for one reaction was composed of: 4.25 μL of water, 1.5 μL of 10 × RT Buffer, 0.15 μL of 100 mM dNTP, 0.1 μL of 40 U/μL RNase Inhibitor (M0307S, New England BioLabs® Inc., Ipswich, MA, USA), 1 μL of 50 U/μL MultiScribe Reverse Transcriptase and 3 μL of 5 × RT-primer specific for each miRNA. .. The RT master mix for one reaction was composed of: 4.25 μL of water, 1.5 μL of 10 × RT Buffer, 0.15 μL of 100 mM dNTP, 0.1 μL of 40 U/μL RNase Inhibitor (M0307S, New England BioLabs® Inc., Ipswich, MA, USA), 1 μL of 50 U/μL MultiScribe Reverse Transcriptase and 3 μL of 5 × RT-primer specific for each miRNA.

    Random Hexamer Labeling:

    Article Title: Lysine-specific Demethylase 2B (KDM2B)-let-
    Article Snippet: Briefly, total RNA, including miRNAs, was isolated either with TRIzol reagent (Invitrogen; 15596-026) or with the mirVana miRNA Isolation Kit (Ambion; AM1560) and dissolved in diethyl pyrocarbonate-treated water. .. Total RNA (1 μg) was polyadenylated using poly(A) polymerase (Ambion; AM2030) according to the manufacturer's instructions and was reverse-transcribed using Superscript III Reverse transcriptase (Invitrogen; 18080-044) in a 20-μl reaction supplemented with 1 μl of random hexamer (New England Biolabs; S1230S), 1 μl of oligo-dT, 1 μl of RNase inhibitor (New England Biolabs; M0307S), and 1 μl of the adapter primer (5′-GCGAGCACAGAATTAATACGACTCACTATAGGTTTTTTTTTTTTVN-3′). .. To quantitate the mature miRNA, a universal reverse primer miRNA-qPCR-3-3′ (5′-GCGAGCACAGAATTAATACGACTCAC-3′) was used in conjunction with an exact sequence-specific primer to each miRNA.

    Gel Extraction:

    Article Title: Quantitative measurement of transcriptional inhibition and mutagenesis induced by site-specifically incorporated DNA lesions in vitro and in vivo
    Article Snippet: Ethanol (Fisher Scientific, cat. no. BP2818) QIAquick gel extraction kit (Qiagen, cat. no. 28704) [γ-32 P]ATP (PerkinElmer, cat. no. NEG002H250UC) ! .. SfaNI (New England BioLabs, cat. no. R0172S) NcoI (New England BioLabs, cat. no. R0193S) NotI (New England BioLabs, cat. no. R0189S) T7 RNA polymerase, supplied with 5× transcription buffer (Promega, cat. no. P2075) 100 mM DTT (Promega, cat. no. P1171) 100 mM rATP (Promega, cat. no. E6011) 100 mM rUTP (Promega, cat. no. E6021) 100 mM rGTP (Promega, cat. no. E6031) 100 mM rCTP (Promega, cat. no. E6041) RNase inhibitor (New England BioLabs, cat. no. M0307S) HeLaScribe nuclear extract in vitro transcription system, supplied with HeLa nuclear extract, 1× transcription buffer and HeLa extract stop solution (Promega, cat. no. E3110) pGEM-T vector (Promega, cat. no. A3600) Lipofectamine 2000 transfection reagent (Invitrogen, cat. no. 11668-019) DMEM (Life Technologies, cat. no. 11995-073) FBS (Life Technologies, cat. no. 16000-044) Penicillin-streptomycin solution (American Type Culture Collection, cat. no. 30-2300) 0.25% (wt/vol) trypsin-EDTA (Life Technologies, cat. no. 25200-056) 10× PBS solution (VWR, cat. no. 97064-158) E.Z.N.A.

    Activity Assay:

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: Paragraph title: Agarose Gel RNase Activity Assay ... For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl.

    Article Title: The vapB–vapC Operon of Acidovorax citrulli Functions as a Bona-fide Toxin–Antitoxin Module
    Article Snippet: Paragraph title: In vitro Analyses of VapC RNase Activity ... The reaction was allowed to proceed for 20 min at room temperature after which it was stopped by addition of 3 μl of 6x DNA loading dye (Thermo Fisher Scientific) and 1 μl of RNase inhibitor (Human Placenta RNase NEB-M0307; 40 units/μl; New England Biolabs).

    Expressing:

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Paragraph title: Localisation of ArLQP expression in A . rubens using mRNA in situ hybridisation ... Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S).

    Modification:

    Article Title: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions
    Article Snippet: PAR-CLIP procedures were performed as previously reported with the following modification. .. The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions.

    Article Title: Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells
    Article Snippet: Next, the cultures were collected 18–24 h after the final transfection, and a modified ChIP assay was performed. .. Next, the cells were washed twice in 1× PBS plus 1/1,000 PMSF (stock PMSF at 0.5M) plus 40U Rnase Inhibitor (New England Biolabs, Ipswich, MA; catalog no. M0307S), resuspended in 600 μl of ChIP lysis buffer (50 mM Hepes, pH 7.5/140 mM NaCl/10% Triton X-100/0.1% NaDeoxycholate/1/1,000 PMSF/40 units RNase inhibitor) and incubated (10 min on ice).

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: RNase assays were performed according to a modified published method. .. For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl.

    Over Expression:

    Article Title: AUTOGENOUS CONTROL OF 5′TOP mRNA STABILITY BY 40S RIBOSOMES
    Article Snippet: The Placental Rnase inhibitor was purchased from NEB (New England Biolabs). .. TetO-MU-L32TOP-β-Globin-MS2(12X)-Puro was generated by site-directed mutagenesis as previously described ( ) using specific primers (see ).

    Hybridization:

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Paragraph title: Localisation of ArLQP expression in A . rubens using mRNA in situ hybridisation ... Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S).

    Article Title: Barriers to transmission of transcriptional noise in a c-fos c-jun pathway
    Article Snippet: To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents. .. To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents.

    Transfection:

    Article Title: Quantitative measurement of transcriptional inhibition and mutagenesis induced by site-specifically incorporated DNA lesions in vitro and in vivo
    Article Snippet: CAUTION [γ-32 P]ATP is radioactive and carcinogenic; use personal protective equipment when you are handling it. .. SfaNI (New England BioLabs, cat. no. R0172S) NcoI (New England BioLabs, cat. no. R0193S) NotI (New England BioLabs, cat. no. R0189S) T7 RNA polymerase, supplied with 5× transcription buffer (Promega, cat. no. P2075) 100 mM DTT (Promega, cat. no. P1171) 100 mM rATP (Promega, cat. no. E6011) 100 mM rUTP (Promega, cat. no. E6021) 100 mM rGTP (Promega, cat. no. E6031) 100 mM rCTP (Promega, cat. no. E6041) RNase inhibitor (New England BioLabs, cat. no. M0307S) HeLaScribe nuclear extract in vitro transcription system, supplied with HeLa nuclear extract, 1× transcription buffer and HeLa extract stop solution (Promega, cat. no. E3110) pGEM-T vector (Promega, cat. no. A3600) Lipofectamine 2000 transfection reagent (Invitrogen, cat. no. 11668-019) DMEM (Life Technologies, cat. no. 11995-073) FBS (Life Technologies, cat. no. 16000-044) Penicillin-streptomycin solution (American Type Culture Collection, cat. no. 30-2300) 0.25% (wt/vol) trypsin-EDTA (Life Technologies, cat. no. 25200-056) 10× PBS solution (VWR, cat. no. 97064-158) E.Z.N.A. .. Total RNA kit I (Omega Bio-Tek, cat. no. 101319-240) Ambion DNA-free kit (Life Technologies, cat. no. AM1906) M-MLV reverse transcriptase, supplied with 5× reaction buffer and HeLa extract stop solution (Promega, cat. no. E3110) MluCI (New England BioLabs, cat. no. R0538S) Cac8I (New England BioLabs, cat. no. R0579L) 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP; Oakwood Chemical, cat. no. 003409) Methanol (Fisher Scientific, cat. no. A452-SK4) Sodium chloride (EMD, cat. no. 7710) Sodium hydroxide (Fisher Scientific, cat. no. S318-10) Agarose (Sigma-Aldrich, cat. no. A9539) Sodium acetate (Sigma-Aldrich, cat. no. S2889) Tris base (Fisher Scientific, cat. no. BP152-5) EDTA (Teknova, cat. no. E5599) Hydrochloric acid (Fisher Scientific, cat. no. A144-500) !

    Article Title: Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells
    Article Snippet: Next, the cultures were collected 18–24 h after the final transfection, and a modified ChIP assay was performed. .. Next, the cells were washed twice in 1× PBS plus 1/1,000 PMSF (stock PMSF at 0.5M) plus 40U Rnase Inhibitor (New England Biolabs, Ipswich, MA; catalog no. M0307S), resuspended in 600 μl of ChIP lysis buffer (50 mM Hepes, pH 7.5/140 mM NaCl/10% Triton X-100/0.1% NaDeoxycholate/1/1,000 PMSF/40 units RNase inhibitor) and incubated (10 min on ice).

    Ligation:

    Article Title: Barriers to transmission of transcriptional noise in a c-fos c-jun pathway
    Article Snippet: To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents. .. To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents.

    Methylation:

    Article Title: Radical SAM-Mediated Methylation of Ribosomal RNA
    Article Snippet: In our earlier investigations, we have determined substrate requirements (i.e., length of the transcribed rRNA fragment) for the methylation of in vitro transcribed rRNA by RlmN and Cfr ( ). .. In a typical in vitro transcription reaction, 20 μg of DNA template is dissolved in 500 μl solution containing T7 buffer (40 m M Tris–HCl, pH 7.9, 16 m M MgCl2 , 2 mM spermidine), 20 m M DTT, 3.2 m M of each of rNTP, 0.2 U/μl RNase inhibitor from human placenta, and 3 U/μl T7 RNA polymerase (New England BioLabs).

    Incubation:

    Article Title: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions
    Article Snippet: PARCLIP-MeRIP experiment applied m6 A-antibody immunoprecipitation , , to the hnRNP C PAR-CLIP RNA samples. .. The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions. .. For each PARCLIP-MeRIP experiment, 20 μl protein-A beads (Invitrogen) were washed twice with 1 ml 1x IP buffer, blocked with 2 hours incubation with 100 μl 1× IP buffer supplemented with BSA (0.5 mg/ml), RNasin and Human placental RNase inhibitor, and then washed twice with 100 μl 1x IP buffer.

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: For inhibition of YoeBSa 1 by YefMSa 1 , YoeBSa 1 (10, 20, or 30 pmol) in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 3 min. YefMSa 1 (0, 10, 20, or 30 pmol) was then added. .. Following incubation at 37°C for 30 min to allow complex formation, 320 ng yefM-yoeB Sa 1 RNA was added with Human Placental RNase Inhibitor at a final concentration of 1 unit/μl. .. For the time course of YoeBSa 1 and YefMSa 1 RNase activity, 10 pmol YoeBSa 1 -Y88ONBY, YoeBSa 1 -Y88F, or YoeBSa 1 -Y88TAG, or YefMSa 1 was incubated with 320 ng yefM-yoeB Sa 1 RNA in the presence of 1 unit/μl Human Placental RNase inhibitor in 50 mM Tris, 5% glycerol, pH 8.0.

    Article Title: RNA Sequences Required for the Noncoding Function of oskar RNA also Mediate Regulation of Oskar Protein Expression by Bicoid Stability Factor
    Article Snippet: 2 μg RNA and 2.5 μg oligo 1 (5'-ggaattcacttgtgactgcg-3') were mixed in 20 μl of 10 mM Tris-HCl, 50 mM NaCl, 1 mM EDTA (pH 8.0), incubated for 10 min at 70 °C and gradually cooled to 25 °C. .. The samples were incubated for 1 hr at 37 °C with 2.5 U of RNase H (NEB) and 20 U of RNase Inhibitor, Human Placenta (HPRI, NEB) in 50 μl of RNase H buffer (NEB). .. Decapped RNAs were purified with RNeasy MiniElute Cleanup Kit (Qiagen).

    Article Title: Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells
    Article Snippet: The ChIP assay consisted of first cross-linking the cultures with formaldehyde [1%, 10 min at room temperature (R/T)] and then the addition of glycine at a final concentration (0.125 M, 10 min at R/T). .. Next, the cells were washed twice in 1× PBS plus 1/1,000 PMSF (stock PMSF at 0.5M) plus 40U Rnase Inhibitor (New England Biolabs, Ipswich, MA; catalog no. M0307S), resuspended in 600 μl of ChIP lysis buffer (50 mM Hepes, pH 7.5/140 mM NaCl/10% Triton X-100/0.1% NaDeoxycholate/1/1,000 PMSF/40 units RNase inhibitor) and incubated (10 min on ice). .. The samples were next centrifuged (5,000 rpm for 5 min at 4°C), resuspended in 600 μl ChIP lysis buffer, incubated (10 min on ice), and then sonicated [Branson 50-cell machine (Branson Ultrasonics Corporation, Danbury, CT), one interval of 20 second constant at an output setting of “3”].

    Article Title: Structural basis for MTR4–ZCCHC8 interactions that stimulate the MTR4 helicase in the nuclear exosome-targeting complex
    Article Snippet: Dissociation constants (K d s) were obtained by fitting the data in the equation: fraction bound = (Bmax [P]h )/(Kd h + [P]h ) where [P] is the protein concentration, h is the Hill’s coefficient, and Bmax is the maximal fraction bound. .. Proteins (200 nM) were incubated with 100 nM RNA (A20 ) labeled with internal 4-thiouridine (4SU) in the indicated position in a buffer containing 20 mM Tris, pH 7.0, 50 mM NaCl, 5 mM BME, 1 U/µL RNase inhibitor, human placenta (New England Biolabs), 1.5 mM MgCl2 , and 1 mM AMPPNP (pH 7.0) at 22 °C for 30 min. Cross-linking was induced by exposure of the protein–RNA mixture to 365 nm UV light using a 4-W handheld UV lamp (UVP) for 20 min at 4 °C. .. Samples were quenched with NuPAGE lithium dodecyl sulfate sample buffer (Thermo Fisher Scientific) and subjected to SDS/PAGE analysis using NuPAGE 4 to 12% Bis-Tris protein gels (Thermo Fisher Scientific).

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl. .. For inhibition of YoeBSa 1 by YefMSa 1 , YoeBSa 1 (10, 20, or 30 pmol) in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 3 min. YefMSa 1 (0, 10, 20, or 30 pmol) was then added.

    Article Title: Barriers to transmission of transcriptional noise in a c-fos c-jun pathway
    Article Snippet: For PLA, we utilized reagents from a Duolink PLA Kit (Olink Biosciences, Uppsala, Sweden). .. To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents. .. For each incubation step, the coverslips were placed over a 50-μl solution (cell-side facing down) on a strip of Parafilm that was stretched over glass and placed in a humid chamber.

    Light Microscopy:

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S). .. Sections of the arms and central disk of A . rubens were prepared and processed for mRNA in situ hybridisation using the same methods reported previously for the neuropeptide precursor ArRGPP .

    Generated:

    Article Title: AUTOGENOUS CONTROL OF 5′TOP mRNA STABILITY BY 40S RIBOSOMES
    Article Snippet: The Placental Rnase inhibitor was purchased from NEB (New England Biolabs). .. The Placental Rnase inhibitor was purchased from NEB (New England Biolabs).

    Inhibition:

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl. .. Reactions were incubated at 37°C for 2 h and stopped by adding 1 μg proteinase K (Invitrogen) and incubating at 37°C for 15 min. 11 μl RNA loading dye I (95% formamide, 18 mM EDTA, 0.025% SDS, 0.025% bromophenol blue) was then added, and samples were incubated at 95°C for 5 min immediately prior to loading on 1.2% agarose, 0.5X TBE (45 mM Tris-borate, 1 mM EDTA, pH 8.3), 0.1 μg/ml EtBr gels for analysis.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: A High-Throughput Screening Assay for the Functional Delivery of Splice-Switching Oligonucleotides in Human Melanoma Cells
    Article Snippet: Paragraph title: 2.3 RT-PCR Assay ... RNase inhibitor, Human Placenta (New England Biolabs). dNTPs (Promega) ( see ).

    Article Title: RNA Sequences Required for the Noncoding Function of oskar RNA also Mediate Regulation of Oskar Protein Expression by Bicoid Stability Factor
    Article Snippet: Paragraph title: RNA circularization and RT-PCR ... The samples were incubated for 1 hr at 37 °C with 2.5 U of RNase H (NEB) and 20 U of RNase Inhibitor, Human Placenta (HPRI, NEB) in 50 μl of RNase H buffer (NEB).

    Sonication:

    Article Title: Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells
    Article Snippet: Next, the cells were washed twice in 1× PBS plus 1/1,000 PMSF (stock PMSF at 0.5M) plus 40U Rnase Inhibitor (New England Biolabs, Ipswich, MA; catalog no. M0307S), resuspended in 600 μl of ChIP lysis buffer (50 mM Hepes, pH 7.5/140 mM NaCl/10% Triton X-100/0.1% NaDeoxycholate/1/1,000 PMSF/40 units RNase inhibitor) and incubated (10 min on ice). .. The samples were next centrifuged (5,000 rpm for 5 min at 4°C), resuspended in 600 μl ChIP lysis buffer, incubated (10 min on ice), and then sonicated [Branson 50-cell machine (Branson Ultrasonics Corporation, Danbury, CT), one interval of 20 second constant at an output setting of “3”].

    Binding Assay:

    Article Title: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions
    Article Snippet: The lysate was then immediately cleared by spinning at 14,000 rpm, 4 °C for 30 min, and placed on ice for further use. hnRNP C binding sites were identified by PARalyzer v1.1 with default settings. .. The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions.

    Pull Down Assay:

    Article Title: Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells
    Article Snippet: Paragraph title: Dual Pull-Down Assay. ... Next, the cells were washed twice in 1× PBS plus 1/1,000 PMSF (stock PMSF at 0.5M) plus 40U Rnase Inhibitor (New England Biolabs, Ipswich, MA; catalog no. M0307S), resuspended in 600 μl of ChIP lysis buffer (50 mM Hepes, pH 7.5/140 mM NaCl/10% Triton X-100/0.1% NaDeoxycholate/1/1,000 PMSF/40 units RNase inhibitor) and incubated (10 min on ice).

    Magnetic Beads:

    Article Title: AUTOGENOUS CONTROL OF 5′TOP mRNA STABILITY BY 40S RIBOSOMES
    Article Snippet: MagnaCHIP Protein A/G magnetic beads mix was from Millipore. .. The Placental Rnase inhibitor was purchased from NEB (New England Biolabs).

    Mutagenesis:

    Article Title: AUTOGENOUS CONTROL OF 5′TOP mRNA STABILITY BY 40S RIBOSOMES
    Article Snippet: The Placental Rnase inhibitor was purchased from NEB (New England Biolabs). .. The Placental Rnase inhibitor was purchased from NEB (New England Biolabs).

    Isolation:

    Article Title: Lysine-specific Demethylase 2B (KDM2B)-let-
    Article Snippet: Paragraph title: Isolation and Quantitation of miRNAs ... Total RNA (1 μg) was polyadenylated using poly(A) polymerase (Ambion; AM2030) according to the manufacturer's instructions and was reverse-transcribed using Superscript III Reverse transcriptase (Invitrogen; 18080-044) in a 20-μl reaction supplemented with 1 μl of random hexamer (New England Biolabs; S1230S), 1 μl of oligo-dT, 1 μl of RNase inhibitor (New England Biolabs; M0307S), and 1 μl of the adapter primer (5′-GCGAGCACAGAATTAATACGACTCACTATAGGTTTTTTTTTTTTVN-3′).

    Article Title: The Evolutionarily Conserved Cassette Exon 7b Drives ERG's Oncogenic Properties
    Article Snippet: Total RNA was extracted using the total RNA isolation mini kit (Agilent Technologies Ltd.). .. All samples were treated with DNAse on the columns using RNase-free DNase I provided in the kit. cDNA was synthesized from 0.2-1 μg of total RNA using 200 U MuLV reverse transcriptase (New England Biolabs), 40 U RNase inhibitor (human placenta) (New England Biolabs), 0.5 mM dNTP, 25 μM oligo-dT primers, and 10× reverse transcriptase buffer (500 mM Tris–HCl pH 8.3, 750 mM KCl, 30 mM MgCl2 , 100 mM DTT) (New England Biolabs) in a final reaction volume 20 μl with added nuclease-free water as required (Qiagen).

    RNA Extraction:

    Article Title: The Evolutionarily Conserved Cassette Exon 7b Drives ERG's Oncogenic Properties
    Article Snippet: Paragraph title: RNA Extraction and cDNA Synthesis ... All samples were treated with DNAse on the columns using RNase-free DNase I provided in the kit. cDNA was synthesized from 0.2-1 μg of total RNA using 200 U MuLV reverse transcriptase (New England Biolabs), 40 U RNase inhibitor (human placenta) (New England Biolabs), 0.5 mM dNTP, 25 μM oligo-dT primers, and 10× reverse transcriptase buffer (500 mM Tris–HCl pH 8.3, 750 mM KCl, 30 mM MgCl2 , 100 mM DTT) (New England Biolabs) in a final reaction volume 20 μl with added nuclease-free water as required (Qiagen).

    Labeling:

    Article Title: Structural basis for MTR4–ZCCHC8 interactions that stimulate the MTR4 helicase in the nuclear exosome-targeting complex
    Article Snippet: Dissociation constants (K d s) were obtained by fitting the data in the equation: fraction bound = (Bmax [P]h )/(Kd h + [P]h ) where [P] is the protein concentration, h is the Hill’s coefficient, and Bmax is the maximal fraction bound. .. Proteins (200 nM) were incubated with 100 nM RNA (A20 ) labeled with internal 4-thiouridine (4SU) in the indicated position in a buffer containing 20 mM Tris, pH 7.0, 50 mM NaCl, 5 mM BME, 1 U/µL RNase inhibitor, human placenta (New England Biolabs), 1.5 mM MgCl2 , and 1 mM AMPPNP (pH 7.0) at 22 °C for 30 min. Cross-linking was induced by exposure of the protein–RNA mixture to 365 nm UV light using a 4-W handheld UV lamp (UVP) for 20 min at 4 °C. .. Samples were quenched with NuPAGE lithium dodecyl sulfate sample buffer (Thermo Fisher Scientific) and subjected to SDS/PAGE analysis using NuPAGE 4 to 12% Bis-Tris protein gels (Thermo Fisher Scientific).

    Purification:

    Article Title: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions
    Article Snippet: The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions. .. The pre-blocked protein-A beads were then combined with the prepared immuno-reaction mixture and incubated at 4 °C for 2 hours, followed by three washes with 100 μl 1× IP buffer.

    Article Title: Radical SAM-Mediated Methylation of Ribosomal RNA
    Article Snippet: The resultant PCR products are purified using the Qiagen PCR purification kit following the manufacturer's recommendations. .. In a typical in vitro transcription reaction, 20 μg of DNA template is dissolved in 500 μl solution containing T7 buffer (40 m M Tris–HCl, pH 7.9, 16 m M MgCl2 , 2 mM spermidine), 20 m M DTT, 3.2 m M of each of rNTP, 0.2 U/μl RNase inhibitor from human placenta, and 3 U/μl T7 RNA polymerase (New England BioLabs).

    Article Title: Primer Extension Reactions for the PCR- based α- complementation Assay
    Article Snippet: .. pBSM13+ (Stratagene) T3 RNA polymerase (Roche Diagnostics, catalog number: P2083) 10x transcription buffer (Roche Diagnostics, catalog number: P2083) High-fidelity PvuII ( PvuII -HF) (New England Biolabs, catalog number: R3151L) High-fidelity EcoRI ( EcoRI -HF) (New England Biolabs, catalog number: R3101L) 10x CutSmart buffer (New England Biolabs, catalog number: R3101L) 6x gel loading dye (New England Biolabs, catalog number: R3101L) NdeI (New England Biolabs, catalog number: R0111L) T4 polynucleotide kinase (PNK) (New England Biolabs, catalog number: M0201L) 10x T4 polynucleotide kinase buffer (New England Biolabs, catalog Number: B0201S) RNasin (RNase inhibitor) (New England Biolabs, catalog number: M0307L) RNase (DNase-free) (Roche Diagnostics, catalog number: 11119915001) RNase-free DNase I (Affymetrix, catalog number: 784111000) Pfu DNA polymerase (Agilent Technologies, catalog number: 600353) 10x Pfu buffer (Agilent Technologies, catalog number: 600353) Ribonucleoside triphosphate set (Roche Diagnostics, catalog number: 11277057001) Deoxynucleoside triphosphate (dNTP) (Roche Diagnostics, catalog number: 11969064001) Gamma [γ-32 P] ATP (PerkinElmer, catalog number: Blu502A001MC) G-25 Macro spin columns (best suited for volumes of 75-150 μl) (Harvard Apparatus, catalog number: 74-3901) RNeasy RNA purification kit (QIAGEN, catalog number: 74104) Phenol: Chloroform: Isoamyl alcohol (25:24:1) (Amresco, catalog number: K169-400ML) Ethanol (VWR Lifesciences, catalog number: EM1.00967.4003) 3M Sodium Acetate (Amresco, catalog number: E521-100ML) Isopropyl alcohol (J.T.Baker® , catalog number: 9037-03) 40% Acrylamide-Bisacrylamide (19:1) solution (VWR International, catalog number: JT4968-0) 40% Acrylamide-Bisacrylamide (29:1) solution (VWR International, catalog number: JT4968-0) Urea (VWR International, catalog number: 97061-926) Ammonium Persulfate (VWR International, catalog number: 97064-594) HIV Reverse Transcriptase, [purified as described in ] Milli-Q quality [RNase, DNase free water (dH2 O)] DNA oligonucleotides were obtained from Integrated DNA Technologies Extension reaction buffer (see Recipes) Elution buffer (see Recipes) 2x SDS loading buffer (see Recipes) .. Eppendorf tubes Micropipette Petri plates Table top centrifuge Incubator Gel apparatus

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Once purified, the DNA was precipitated by adding 1/10 volume of 3 M sodium acetate and 2.5 volumes of 100% isopropanol and incubating at −80 °C for 30 min. .. Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S).

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: RNA was purified with reagents from the Total RNA Kit I (Omega Bio-Tek) according to the RNA Cleanup protocol from the RNeasy Mini Handbook (Qiagen). .. For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl.

    Polymerase Chain Reaction:

    Article Title: Radical SAM-Mediated Methylation of Ribosomal RNA
    Article Snippet: The resultant PCR products are purified using the Qiagen PCR purification kit following the manufacturer's recommendations. .. In a typical in vitro transcription reaction, 20 μg of DNA template is dissolved in 500 μl solution containing T7 buffer (40 m M Tris–HCl, pH 7.9, 16 m M MgCl2 , 2 mM spermidine), 20 m M DTT, 3.2 m M of each of rNTP, 0.2 U/μl RNase inhibitor from human placenta, and 3 U/μl T7 RNA polymerase (New England BioLabs).

    Article Title: A High-Throughput Screening Assay for the Functional Delivery of Splice-Switching Oligonucleotides in Human Melanoma Cells
    Article Snippet: RNase inhibitor, Human Placenta (New England Biolabs). dNTPs (Promega) ( see ). .. RNase inhibitor, Human Placenta (New England Biolabs). dNTPs (Promega) ( see ).

    Article Title: RNA Sequences Required for the Noncoding Function of oskar RNA also Mediate Regulation of Oskar Protein Expression by Bicoid Stability Factor
    Article Snippet: The samples were incubated for 1 hr at 37 °C with 2.5 U of RNase H (NEB) and 20 U of RNase Inhibitor, Human Placenta (HPRI, NEB) in 50 μl of RNase H buffer (NEB). .. The samples were incubated for 1 hr at 37 °C with 2.5 U of RNase H (NEB) and 20 U of RNase Inhibitor, Human Placenta (HPRI, NEB) in 50 μl of RNase H buffer (NEB).

    Article Title: Stabilization of Urinary MicroRNAs by Association with Exosomes and Argonaute 2 Protein
    Article Snippet: Paragraph title: 5.3. Reverse Transcription-Quantitative PCR (RT-qPCR) Analysis ... The RT master mix for one reaction was composed of: 4.25 μL of water, 1.5 μL of 10 × RT Buffer, 0.15 μL of 100 mM dNTP, 0.1 μL of 40 U/μL RNase Inhibitor (M0307S, New England BioLabs® Inc., Ipswich, MA, USA), 1 μL of 50 U/μL MultiScribe Reverse Transcriptase and 3 μL of 5 × RT-primer specific for each miRNA.

    In Vitro:

    Article Title: Radical SAM-Mediated Methylation of Ribosomal RNA
    Article Snippet: The resultant PCR products are purified using the Qiagen PCR purification kit following the manufacturer's recommendations. .. In a typical in vitro transcription reaction, 20 μg of DNA template is dissolved in 500 μl solution containing T7 buffer (40 m M Tris–HCl, pH 7.9, 16 m M MgCl2 , 2 mM spermidine), 20 m M DTT, 3.2 m M of each of rNTP, 0.2 U/μl RNase inhibitor from human placenta, and 3 U/μl T7 RNA polymerase (New England BioLabs). .. The reaction mixture is incubated at 37 °C for 3 h, followed by the addition of 40 U of RQ1 RNase Free DNase (Promega) and further incubation at 37 °C for 30 min.

    Article Title: Quantitative measurement of transcriptional inhibition and mutagenesis induced by site-specifically incorporated DNA lesions in vitro and in vivo
    Article Snippet: CAUTION [γ-32 P]ATP is radioactive and carcinogenic; use personal protective equipment when you are handling it. .. SfaNI (New England BioLabs, cat. no. R0172S) NcoI (New England BioLabs, cat. no. R0193S) NotI (New England BioLabs, cat. no. R0189S) T7 RNA polymerase, supplied with 5× transcription buffer (Promega, cat. no. P2075) 100 mM DTT (Promega, cat. no. P1171) 100 mM rATP (Promega, cat. no. E6011) 100 mM rUTP (Promega, cat. no. E6021) 100 mM rGTP (Promega, cat. no. E6031) 100 mM rCTP (Promega, cat. no. E6041) RNase inhibitor (New England BioLabs, cat. no. M0307S) HeLaScribe nuclear extract in vitro transcription system, supplied with HeLa nuclear extract, 1× transcription buffer and HeLa extract stop solution (Promega, cat. no. E3110) pGEM-T vector (Promega, cat. no. A3600) Lipofectamine 2000 transfection reagent (Invitrogen, cat. no. 11668-019) DMEM (Life Technologies, cat. no. 11995-073) FBS (Life Technologies, cat. no. 16000-044) Penicillin-streptomycin solution (American Type Culture Collection, cat. no. 30-2300) 0.25% (wt/vol) trypsin-EDTA (Life Technologies, cat. no. 25200-056) 10× PBS solution (VWR, cat. no. 97064-158) E.Z.N.A. .. Total RNA kit I (Omega Bio-Tek, cat. no. 101319-240) Ambion DNA-free kit (Life Technologies, cat. no. AM1906) M-MLV reverse transcriptase, supplied with 5× reaction buffer and HeLa extract stop solution (Promega, cat. no. E3110) MluCI (New England BioLabs, cat. no. R0538S) Cac8I (New England BioLabs, cat. no. R0579L) 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP; Oakwood Chemical, cat. no. 003409) Methanol (Fisher Scientific, cat. no. A452-SK4) Sodium chloride (EMD, cat. no. 7710) Sodium hydroxide (Fisher Scientific, cat. no. S318-10) Agarose (Sigma-Aldrich, cat. no. A9539) Sodium acetate (Sigma-Aldrich, cat. no. S2889) Tris base (Fisher Scientific, cat. no. BP152-5) EDTA (Teknova, cat. no. E5599) Hydrochloric acid (Fisher Scientific, cat. no. A144-500) !

    Article Title: The vapB–vapC Operon of Acidovorax citrulli Functions as a Bona-fide Toxin–Antitoxin Module
    Article Snippet: Paragraph title: In vitro Analyses of VapC RNase Activity ... The reaction was allowed to proceed for 20 min at room temperature after which it was stopped by addition of 3 μl of 6x DNA loading dye (Thermo Fisher Scientific) and 1 μl of RNase inhibitor (Human Placenta RNase NEB-M0307; 40 units/μl; New England Biolabs).

    Positron Emission Tomography:

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: pET-28a- yefM-yoeB Sa 1 was digested with XhoI (NEB) at 37°C for 3 h. The fully linearized plasmid was extracted once with 24.5:24.5:1 phenol:chloroform:isoamyl alcohol and twice with chloroform, precipitated with 3 M NaOAc, pH 5.3, and 100% EtOH, and resuspended in nuclease-free water. .. For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl.

    Blocking Assay:

    Article Title: Barriers to transmission of transcriptional noise in a c-fos c-jun pathway
    Article Snippet: To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents. .. To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents.

    Lysis:

    Article Title: Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells
    Article Snippet: The ChIP assay consisted of first cross-linking the cultures with formaldehyde [1%, 10 min at room temperature (R/T)] and then the addition of glycine at a final concentration (0.125 M, 10 min at R/T). .. Next, the cells were washed twice in 1× PBS plus 1/1,000 PMSF (stock PMSF at 0.5M) plus 40U Rnase Inhibitor (New England Biolabs, Ipswich, MA; catalog no. M0307S), resuspended in 600 μl of ChIP lysis buffer (50 mM Hepes, pH 7.5/140 mM NaCl/10% Triton X-100/0.1% NaDeoxycholate/1/1,000 PMSF/40 units RNase inhibitor) and incubated (10 min on ice). .. The samples were next centrifuged (5,000 rpm for 5 min at 4°C), resuspended in 600 μl ChIP lysis buffer, incubated (10 min on ice), and then sonicated [Branson 50-cell machine (Branson Ultrasonics Corporation, Danbury, CT), one interval of 20 second constant at an output setting of “3”].

    Concentration Assay:

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: For inhibition of YoeBSa 1 by YefMSa 1 , YoeBSa 1 (10, 20, or 30 pmol) in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 3 min. YefMSa 1 (0, 10, 20, or 30 pmol) was then added. .. Following incubation at 37°C for 30 min to allow complex formation, 320 ng yefM-yoeB Sa 1 RNA was added with Human Placental RNase Inhibitor at a final concentration of 1 unit/μl. .. For the time course of YoeBSa 1 and YefMSa 1 RNase activity, 10 pmol YoeBSa 1 -Y88ONBY, YoeBSa 1 -Y88F, or YoeBSa 1 -Y88TAG, or YefMSa 1 was incubated with 320 ng yefM-yoeB Sa 1 RNA in the presence of 1 unit/μl Human Placental RNase inhibitor in 50 mM Tris, 5% glycerol, pH 8.0.

    Article Title: Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells
    Article Snippet: The ChIP assay consisted of first cross-linking the cultures with formaldehyde [1%, 10 min at room temperature (R/T)] and then the addition of glycine at a final concentration (0.125 M, 10 min at R/T). .. Next, the cells were washed twice in 1× PBS plus 1/1,000 PMSF (stock PMSF at 0.5M) plus 40U Rnase Inhibitor (New England Biolabs, Ipswich, MA; catalog no. M0307S), resuspended in 600 μl of ChIP lysis buffer (50 mM Hepes, pH 7.5/140 mM NaCl/10% Triton X-100/0.1% NaDeoxycholate/1/1,000 PMSF/40 units RNase inhibitor) and incubated (10 min on ice).

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: RNase assays were performed according to a modified published method. .. For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl. .. Reactions were incubated at 37°C for 2 h and stopped by adding 1 μg proteinase K (Invitrogen) and incubating at 37°C for 15 min. 11 μl RNA loading dye I (95% formamide, 18 mM EDTA, 0.025% SDS, 0.025% bromophenol blue) was then added, and samples were incubated at 95°C for 5 min immediately prior to loading on 1.2% agarose, 0.5X TBE (45 mM Tris-borate, 1 mM EDTA, pH 8.3), 0.1 μg/ml EtBr gels for analysis.

    Sample Prep:

    Article Title: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions
    Article Snippet: The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions. .. The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions.

    Chromatin Immunoprecipitation:

    Article Title: Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells
    Article Snippet: The ChIP assay consisted of first cross-linking the cultures with formaldehyde [1%, 10 min at room temperature (R/T)] and then the addition of glycine at a final concentration (0.125 M, 10 min at R/T). .. Next, the cells were washed twice in 1× PBS plus 1/1,000 PMSF (stock PMSF at 0.5M) plus 40U Rnase Inhibitor (New England Biolabs, Ipswich, MA; catalog no. M0307S), resuspended in 600 μl of ChIP lysis buffer (50 mM Hepes, pH 7.5/140 mM NaCl/10% Triton X-100/0.1% NaDeoxycholate/1/1,000 PMSF/40 units RNase inhibitor) and incubated (10 min on ice). .. The samples were next centrifuged (5,000 rpm for 5 min at 4°C), resuspended in 600 μl ChIP lysis buffer, incubated (10 min on ice), and then sonicated [Branson 50-cell machine (Branson Ultrasonics Corporation, Danbury, CT), one interval of 20 second constant at an output setting of “3”].

    Plasmid Preparation:

    Article Title: Radical SAM-Mediated Methylation of Ribosomal RNA
    Article Snippet: In vitro transcription reactions of 23S rRNA fragments are carried out in the following manner: in the first step, a desired fragment of the DNA template is amplified using a plasmid pKK3535 that encodes a 23S rRNA gene ( ). .. In a typical in vitro transcription reaction, 20 μg of DNA template is dissolved in 500 μl solution containing T7 buffer (40 m M Tris–HCl, pH 7.9, 16 m M MgCl2 , 2 mM spermidine), 20 m M DTT, 3.2 m M of each of rNTP, 0.2 U/μl RNase inhibitor from human placenta, and 3 U/μl T7 RNA polymerase (New England BioLabs).

    Article Title: Quantitative measurement of transcriptional inhibition and mutagenesis induced by site-specifically incorporated DNA lesions in vitro and in vivo
    Article Snippet: CAUTION [γ-32 P]ATP is radioactive and carcinogenic; use personal protective equipment when you are handling it. .. SfaNI (New England BioLabs, cat. no. R0172S) NcoI (New England BioLabs, cat. no. R0193S) NotI (New England BioLabs, cat. no. R0189S) T7 RNA polymerase, supplied with 5× transcription buffer (Promega, cat. no. P2075) 100 mM DTT (Promega, cat. no. P1171) 100 mM rATP (Promega, cat. no. E6011) 100 mM rUTP (Promega, cat. no. E6021) 100 mM rGTP (Promega, cat. no. E6031) 100 mM rCTP (Promega, cat. no. E6041) RNase inhibitor (New England BioLabs, cat. no. M0307S) HeLaScribe nuclear extract in vitro transcription system, supplied with HeLa nuclear extract, 1× transcription buffer and HeLa extract stop solution (Promega, cat. no. E3110) pGEM-T vector (Promega, cat. no. A3600) Lipofectamine 2000 transfection reagent (Invitrogen, cat. no. 11668-019) DMEM (Life Technologies, cat. no. 11995-073) FBS (Life Technologies, cat. no. 16000-044) Penicillin-streptomycin solution (American Type Culture Collection, cat. no. 30-2300) 0.25% (wt/vol) trypsin-EDTA (Life Technologies, cat. no. 25200-056) 10× PBS solution (VWR, cat. no. 97064-158) E.Z.N.A. .. Total RNA kit I (Omega Bio-Tek, cat. no. 101319-240) Ambion DNA-free kit (Life Technologies, cat. no. AM1906) M-MLV reverse transcriptase, supplied with 5× reaction buffer and HeLa extract stop solution (Promega, cat. no. E3110) MluCI (New England BioLabs, cat. no. R0538S) Cac8I (New England BioLabs, cat. no. R0579L) 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP; Oakwood Chemical, cat. no. 003409) Methanol (Fisher Scientific, cat. no. A452-SK4) Sodium chloride (EMD, cat. no. 7710) Sodium hydroxide (Fisher Scientific, cat. no. S318-10) Agarose (Sigma-Aldrich, cat. no. A9539) Sodium acetate (Sigma-Aldrich, cat. no. S2889) Tris base (Fisher Scientific, cat. no. BP152-5) EDTA (Teknova, cat. no. E5599) Hydrochloric acid (Fisher Scientific, cat. no. A144-500) !

    Article Title: AUTOGENOUS CONTROL OF 5′TOP mRNA STABILITY BY 40S RIBOSOMES
    Article Snippet: The Placental Rnase inhibitor was purchased from NEB (New England Biolabs). .. TetO-MU-L32TOP-β-Globin-MS2(12X)-Puro was generated by site-directed mutagenesis as previously described ( ) using specific primers (see ).

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Following centrifugation for 10 min at 13,000 rpm in an Eppendorf 5424 R centrifuge (Eppendorf; Cat. No. 5404000138) at 4 °C, the precipitated DNA was washed with 70% ice-cold ethanol before air drying and re-suspending in RNAse-free water. .. Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S). .. The mixture was incubated for 2 hours at 37 °C to allow the in vitro transcription.

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: 1 μg linearized plasmid was used as template for standard RNA synthesis with the T7 High Yield RNA Synthesis Kit (NEB) according to the manufacturer’s instructions. .. For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl.

    Software:

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S). .. Sections of the arms and central disk of A . rubens were prepared and processed for mRNA in situ hybridisation using the same methods reported previously for the neuropeptide precursor ArRGPP .

    Negative Control:

    Article Title: Stabilization of Urinary MicroRNAs by Association with Exosomes and Argonaute 2 Protein
    Article Snippet: The RT master mix for one reaction was composed of: 4.25 μL of water, 1.5 μL of 10 × RT Buffer, 0.15 μL of 100 mM dNTP, 0.1 μL of 40 U/μL RNase Inhibitor (M0307S, New England BioLabs® Inc., Ipswich, MA, USA), 1 μL of 50 U/μL MultiScribe Reverse Transcriptase and 3 μL of 5 × RT-primer specific for each miRNA. .. The RT master mix for one reaction was composed of: 4.25 μL of water, 1.5 μL of 10 × RT Buffer, 0.15 μL of 100 mM dNTP, 0.1 μL of 40 U/μL RNase Inhibitor (M0307S, New England BioLabs® Inc., Ipswich, MA, USA), 1 μL of 50 U/μL MultiScribe Reverse Transcriptase and 3 μL of 5 × RT-primer specific for each miRNA.

    Recombinant:

    Article Title: The vapB–vapC Operon of Acidovorax citrulli Functions as a Bona-fide Toxin–Antitoxin Module
    Article Snippet: One microgram of total RNA from A. citrulli 7a1 was added to 1 μg of refolded VapC-6xHis recombinant protein in 20 μl of reaction buffer containing 50 mM Tris-HCl and 6 mM MgCl (pH 7). .. The reaction was allowed to proceed for 20 min at room temperature after which it was stopped by addition of 3 μl of 6x DNA loading dye (Thermo Fisher Scientific) and 1 μl of RNase inhibitor (Human Placenta RNase NEB-M0307; 40 units/μl; New England Biolabs).

    Agarose Gel Electrophoresis:

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity
    Article Snippet: Paragraph title: Agarose Gel RNase Activity Assay ... For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl.

    In Situ:

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Paragraph title: Localisation of ArLQP expression in A . rubens using mRNA in situ hybridisation ... Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S).

    Article Title: Barriers to transmission of transcriptional noise in a c-fos c-jun pathway
    Article Snippet: To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents. .. To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents.

    Proximity Ligation Assay:

    Article Title: Barriers to transmission of transcriptional noise in a c-fos c-jun pathway
    Article Snippet: For PLA, we utilized reagents from a Duolink PLA Kit (Olink Biosciences, Uppsala, Sweden). .. To protect the cellular RNA during PLA, we included RNase inhibitor from human placenta (New England Biolabs) in the incubation steps, included ribonucleoside-vanadyl complex in washing buffers, and used diethylpyrocarbonate-treated water for dilution of the reagents. .. For each incubation step, the coverslips were placed over a 50-μl solution (cell-side facing down) on a strip of Parafilm that was stretched over glass and placed in a humid chamber.

    Ethanol Precipitation:

    Article Title: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions
    Article Snippet: The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions. .. The pre-blocked protein-A beads were then combined with the prepared immuno-reaction mixture and incubated at 4 °C for 2 hours, followed by three washes with 100 μl 1× IP buffer.

    Quantitation Assay:

    Article Title: Lysine-specific Demethylase 2B (KDM2B)-let-
    Article Snippet: Paragraph title: Isolation and Quantitation of miRNAs ... Total RNA (1 μg) was polyadenylated using poly(A) polymerase (Ambion; AM2030) according to the manufacturer's instructions and was reverse-transcribed using Superscript III Reverse transcriptase (Invitrogen; 18080-044) in a 20-μl reaction supplemented with 1 μl of random hexamer (New England Biolabs; S1230S), 1 μl of oligo-dT, 1 μl of RNase inhibitor (New England Biolabs; M0307S), and 1 μl of the adapter primer (5′-GCGAGCACAGAATTAATACGACTCACTATAGGTTTTTTTTTTTTVN-3′).

    Immunoprecipitation:

    Article Title: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions
    Article Snippet: PARCLIP-MeRIP experiment applied m6 A-antibody immunoprecipitation , , to the hnRNP C PAR-CLIP RNA samples. .. The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions.

    Staining:

    Article Title: Discovery and functional characterisation of a luqin-type neuropeptide signalling system in a deuterostome
    Article Snippet: Sense and antisense RNA probes were synthesised using 1 μg of the previously linearised plasmid DNA as a template, 0.5 mM digoxigenin RNA labelling mix (Roche, Basel, Switzerland, Cat. No. 11277073910), 1 × transcription buffer (NEB, Ipswich, USA, Cat. No. M0251S), 5 mM dithiothreitol (Promega, Madison, USA, Cat. No. P1171), 1 U/μl placental ribonuclease inhibitor (NEB, Ipswich, USA, Cat. No. M0307S) and 5 U/μl of T7 RNA polymerase (NEB, Ipswich, USA, Cat. No. M0251S) for the antisense probe or 5 U/μl of SP6 RNA polymerase for the sense probe (NEB, Ipswich, USA, Cat. No. M0207S). .. Sections of the arms and central disk of A . rubens were prepared and processed for mRNA in situ hybridisation using the same methods reported previously for the neuropeptide precursor ArRGPP .

    Article Title: The vapB–vapC Operon of Acidovorax citrulli Functions as a Bona-fide Toxin–Antitoxin Module
    Article Snippet: The reaction was allowed to proceed for 20 min at room temperature after which it was stopped by addition of 3 μl of 6x DNA loading dye (Thermo Fisher Scientific) and 1 μl of RNase inhibitor (Human Placenta RNase NEB-M0307; 40 units/μl; New England Biolabs). .. The reaction was allowed to proceed for 20 min at room temperature after which it was stopped by addition of 3 μl of 6x DNA loading dye (Thermo Fisher Scientific) and 1 μl of RNase inhibitor (Human Placenta RNase NEB-M0307; 40 units/μl; New England Biolabs).

    Hood:

    Article Title: Quantitative measurement of transcriptional inhibition and mutagenesis induced by site-specifically incorporated DNA lesions in vitro and in vivo
    Article Snippet: SfaNI (New England BioLabs, cat. no. R0172S) NcoI (New England BioLabs, cat. no. R0193S) NotI (New England BioLabs, cat. no. R0189S) T7 RNA polymerase, supplied with 5× transcription buffer (Promega, cat. no. P2075) 100 mM DTT (Promega, cat. no. P1171) 100 mM rATP (Promega, cat. no. E6011) 100 mM rUTP (Promega, cat. no. E6021) 100 mM rGTP (Promega, cat. no. E6031) 100 mM rCTP (Promega, cat. no. E6041) RNase inhibitor (New England BioLabs, cat. no. M0307S) HeLaScribe nuclear extract in vitro transcription system, supplied with HeLa nuclear extract, 1× transcription buffer and HeLa extract stop solution (Promega, cat. no. E3110) pGEM-T vector (Promega, cat. no. A3600) Lipofectamine 2000 transfection reagent (Invitrogen, cat. no. 11668-019) DMEM (Life Technologies, cat. no. 11995-073) FBS (Life Technologies, cat. no. 16000-044) Penicillin-streptomycin solution (American Type Culture Collection, cat. no. 30-2300) 0.25% (wt/vol) trypsin-EDTA (Life Technologies, cat. no. 25200-056) 10× PBS solution (VWR, cat. no. 97064-158) E.Z.N.A. .. SfaNI (New England BioLabs, cat. no. R0172S) NcoI (New England BioLabs, cat. no. R0193S) NotI (New England BioLabs, cat. no. R0189S) T7 RNA polymerase, supplied with 5× transcription buffer (Promega, cat. no. P2075) 100 mM DTT (Promega, cat. no. P1171) 100 mM rATP (Promega, cat. no. E6011) 100 mM rUTP (Promega, cat. no. E6021) 100 mM rGTP (Promega, cat. no. E6031) 100 mM rCTP (Promega, cat. no. E6041) RNase inhibitor (New England BioLabs, cat. no. M0307S) HeLaScribe nuclear extract in vitro transcription system, supplied with HeLa nuclear extract, 1× transcription buffer and HeLa extract stop solution (Promega, cat. no. E3110) pGEM-T vector (Promega, cat. no. A3600) Lipofectamine 2000 transfection reagent (Invitrogen, cat. no. 11668-019) DMEM (Life Technologies, cat. no. 11995-073) FBS (Life Technologies, cat. no. 16000-044) Penicillin-streptomycin solution (American Type Culture Collection, cat. no. 30-2300) 0.25% (wt/vol) trypsin-EDTA (Life Technologies, cat. no. 25200-056) 10× PBS solution (VWR, cat. no. 97064-158) E.Z.N.A.

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    New England Biolabs human placental rnase inhibitor
    <t>RNase</t> activity of <t>YoeB</t> Sa 1 mutants
    Human Placental Rnase Inhibitor, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    RNase activity of YoeB Sa 1 mutants

    Journal:

    Article Title: Light Activation of Staphylococcus aureus Toxin YoeBSa1 Reveals Guanosine-Specific Endoribonuclease Activity

    doi: 10.1021/bi4008098

    Figure Lengend Snippet: RNase activity of YoeB Sa 1 mutants

    Article Snippet: For light-dependence of decaging, 10 pmol YoeBSa 1 -Y88ONBY in 50 mM Tris, 5% glycerol, pH 8.0, was exposed to UV light for 0–5 min. 320 ng yefM-yoeB Sa 1 RNA was then added with Human Placental RNase Inhibitor (NEB) at a final concentration of 1 unit/μl.

    Techniques: Activity Assay