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:
    323
    Size:
    10 000 units
    Category:
    Enzyme Inhibitors
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    New England Biolabs rnase inhibitor
    RNase Inhibitor Human Placenta
    RNase Inhibitor Human Placenta 10 000 units
    https://www.bioz.com/result/rnase inhibitor/product/New England Biolabs
    Average 95 stars, based on 48 article reviews
    Price from $9.99 to $1999.99
    rnase inhibitor - by Bioz Stars, 2020-02
    95/100 stars

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    Related Articles

    Centrifugation:

    Article Title: MitoRibo-Tag Mice Provide a Tool for In Vivo Studies of Mitoribosome Composition
    Article Snippet: Mitochondria (1 mg from mouse tissue or 750 μg from cultured cells) were re-isolated by centrifugation for 5 min/9200 g/4°C. .. Mitochondria were resuspended at protein concentrations of 7.5 mg/ml (from cultured cells) or 10 mg/ml (from mouse tissues) in lysis buffer (260 mM sucrose, 10 mM Tris-HCl pH 7.5, 100 mM KCl, 20 mM MgCl2 , 2% digitonin for mouse mitochondria or 6% digitonin for human mitochondria, 40 U/ml RNase inhibitor (New England Biolabs, cat. no. M0307L) and 1x PIC) and incubated for 20 min on ice.

    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: Circulating and Urinary miR-210 and miR-16 Increase during Cardiac Surgery Using Cardiopulmonary Bypass – A Pilot Study
    Article Snippet: After extraction of RNA, 10 μL of sample was pretreated with .5 μL of the RNase Inhibitor (Cat No M0307L, New England BioLabs® Inc, Ipswich, MA) and 2 units of Heparinase I (Cat No H2519, Sigma-Aldrich® , St Louis, MO) [in 20 mM Tris-HCl {pH 7.5}, 50 mM NaCl, 4 mM CaCl2 ] for 1 hour at 25°C to remove heparin ( ). .. Preliminary experiments showed that heparinase does not affect polymerase chain reaction (PCR) analyses in serum and urine from non-heparinized patients, while allowing unaffected PCR amplification in samples potentially contaminated with heparin.

    Random Hexamer Labeling:

    Article Title: Lysine-specific Demethylase 2B (KDM2B)-let-7-Enhancer of Zester Homolog 2 (EZH2) Pathway Regulates Cell Cycle Progression and Senescence in Primary Cells *
    Article Snippet: .. 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.

    Synthesized:

    Article Title: Detection of Endogenous MazF Enzymatic Activity in Staphylococcus aureus
    Article Snippet: Primers and oligonucleotides were synthesized by IDT. .. Restriction enzymes, BSA, Low-Range ssRNA Ladder, RNase Inhibitor—Human Placenta, and E. coli strains DH5α and NiCo21(DE3) [ ] were purchased from New England Biolabs (NEB).

    Article Title: The Evolutionarily Conserved Cassette Exon 7b Drives ERG's Oncogenic Properties
    Article Snippet: .. 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: NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells
    Article Snippet: .. RT-PCR/qPCR RNA was isolated from canine expanded cells using RNAeasy Kit, QiaShredder Columns, and RNAase-Free DNase Set (all Qiagen, 74104, 79654, 79254), and cDNA was synthesized using Omniscript RT Kit (Qiagen, 20511) with RNase inhibitor (New England Biolabs, M0307S) and Oligo(dt)20 (ThermoFisher Scientific, 18418-020). .. RT-PCR and qPCR reactions were performed on Roche 480 (Roche, USA) using Power Up Sybr Green Master Mix (ThermoFisher; A25742) or TaqMan Universal PCR Mastermix for qPCR for CD16 (ThermoFisher Scientific, 4304437) for 40 cycles.

    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: NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells
    Article Snippet: RT-PCR/qPCR RNA was isolated from canine expanded cells using RNAeasy Kit, QiaShredder Columns, and RNAase-Free DNase Set (all Qiagen, 74104, 79654, 79254), and cDNA was synthesized using Omniscript RT Kit (Qiagen, 20511) with RNase inhibitor (New England Biolabs, M0307S) and Oligo(dt)20 (ThermoFisher Scientific, 18418-020). .. RT-PCR and qPCR reactions were performed on Roche 480 (Roche, USA) using Power Up Sybr Green Master Mix (ThermoFisher; A25742) or TaqMan Universal PCR Mastermix for qPCR for CD16 (ThermoFisher Scientific, 4304437) for 40 cycles.

    Incubation:

    Article Title: Knockdown of fucosyltransferase III disrupts the adhesion of circulating cancer cells to E-selectin without affecting hematopoietic cell adhesion
    Article Snippet: The 40-μL reverse transcription reaction system included 5–10 μg of total RNA, 1 μL of M-MuLV Reverse Transcriptase (Cat# M0253L, New England Biolabs), 0.5 μL of RNase Inhibitor (Cat# M0307S, New England Biolabs), 1 μL of Random Primers (Cat# 48190-011, Invitrogen), 2.5 μL of dNTP Mix (Cat# 170-8874, New England Biolabs) and 4 μL of M-MuLV Reverse Transcriptase reaction buffer. .. The reaction mixture was incubated at 42 °C for 1 h followed by an inactivation step at 95 °C for 10 min.

    Article Title: MitoRibo-Tag Mice Provide a Tool for In Vivo Studies of Mitoribosome Composition
    Article Snippet: .. Mitochondria were resuspended at protein concentrations of 7.5 mg/ml (from cultured cells) or 10 mg/ml (from mouse tissues) in lysis buffer (260 mM sucrose, 10 mM Tris-HCl pH 7.5, 100 mM KCl, 20 mM MgCl2 , 2% digitonin for mouse mitochondria or 6% digitonin for human mitochondria, 40 U/ml RNase inhibitor (New England Biolabs, cat. no. M0307L) and 1x PIC) and incubated for 20 min on ice. .. Samples were clarified by centrifugation for 45 min/9200 g/4°C.

    Article Title: Data on the inhibition of RNase inhibitor activity by a monoclonal antibody as assessed by microfluidics-based RNA electrophoresis
    Article Snippet: 2.2 Inhibition of RI activity by a monoclonal Ab specific to RI (3F11) Purified total RNA from Vero cells (5 µg in 200 µL of CL Buffer) was mixed with 1 µg (1 µL) of monoclonal Ab specific to either RI (Origene; TA501875; clone 3F11) or GAPDH (Origene; TA802519; clone 2D9), 1 ng (1 µL) of RNase A (Qiagen; 19101; diluted in CL Buffer), 40 units (1 µL) of human placental RI (hpRI; New England Biolabs; M0307), or a combination of the above; in reactions containing hpRI, its addition preceded the addition of other components. .. Following the incubation, RNA was purified and subjected to Experion analysis.

    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). .. The mixture was incubated for 2 hours at 37 °C to allow the in vitro transcription.

    Activity Assay:

    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).

    Article Title: Data on the inhibition of RNase inhibitor activity by a monoclonal antibody as assessed by microfluidics-based RNA electrophoresis
    Article Snippet: .. 2.2 Inhibition of RI activity by a monoclonal Ab specific to RI (3F11) Purified total RNA from Vero cells (5 µg in 200 µL of CL Buffer) was mixed with 1 µg (1 µL) of monoclonal Ab specific to either RI (Origene; TA501875; clone 3F11) or GAPDH (Origene; TA802519; clone 2D9), 1 ng (1 µL) of RNase A (Qiagen; 19101; diluted in CL Buffer), 40 units (1 µL) of human placental RI (hpRI; New England Biolabs; M0307), or a combination of the above; in reactions containing hpRI, its addition preceded the addition of other components. .. Following the incubation, RNA was purified and subjected to Experion analysis.

    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).

    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).

    Transfection:

    Article Title: Knockdown of fucosyltransferase III disrupts the adhesion of circulating cancer cells to E-selectin without affecting hematopoietic cell adhesion
    Article Snippet: Four days after siRNA transfection, total RNA was purified using RNeasy Plus mini Kit (Cat# 74134 Qiagen). .. The 40-μL reverse transcription reaction system included 5–10 μg of total RNA, 1 μL of M-MuLV Reverse Transcriptase (Cat# M0253L, New England Biolabs), 0.5 μL of RNase Inhibitor (Cat# M0307S, New England Biolabs), 1 μL of Random Primers (Cat# 48190-011, Invitrogen), 2.5 μL of dNTP Mix (Cat# 170-8874, New England Biolabs) and 4 μL of M-MuLV Reverse Transcriptase reaction buffer.

    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. .. 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) !

    Sequencing:

    Article Title: Lysine-specific Demethylase 2B (KDM2B)-let-7-Enhancer of Zester Homolog 2 (EZH2) Pathway Regulates Cell Cycle Progression and Senescence in Primary Cells *
    Article Snippet: 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.

    Cell Culture:

    Article Title: MitoRibo-Tag Mice Provide a Tool for In Vivo Studies of Mitoribosome Composition
    Article Snippet: .. Mitochondria were resuspended at protein concentrations of 7.5 mg/ml (from cultured cells) or 10 mg/ml (from mouse tissues) in lysis buffer (260 mM sucrose, 10 mM Tris-HCl pH 7.5, 100 mM KCl, 20 mM MgCl2 , 2% digitonin for mouse mitochondria or 6% digitonin for human mitochondria, 40 U/ml RNase inhibitor (New England Biolabs, cat. no. M0307L) and 1x PIC) and incubated for 20 min on ice. .. Samples were clarified by centrifugation for 45 min/9200 g/4°C.

    Inhibition:

    Article Title: Data on the inhibition of RNase inhibitor activity by a monoclonal antibody as assessed by microfluidics-based RNA electrophoresis
    Article Snippet: .. 2.2 Inhibition of RI activity by a monoclonal Ab specific to RI (3F11) Purified total RNA from Vero cells (5 µg in 200 µL of CL Buffer) was mixed with 1 µg (1 µL) of monoclonal Ab specific to either RI (Origene; TA501875; clone 3F11) or GAPDH (Origene; TA802519; clone 2D9), 1 ng (1 µL) of RNase A (Qiagen; 19101; diluted in CL Buffer), 40 units (1 µL) of human placental RI (hpRI; New England Biolabs; M0307), or a combination of the above; in reactions containing hpRI, its addition preceded the addition of other components. .. Following the incubation, RNA was purified and subjected to Experion analysis.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells
    Article Snippet: RT-PCR/qPCR RNA was isolated from canine expanded cells using RNAeasy Kit, QiaShredder Columns, and RNAase-Free DNase Set (all Qiagen, 74104, 79654, 79254), and cDNA was synthesized using Omniscript RT Kit (Qiagen, 20511) with RNase inhibitor (New England Biolabs, M0307S) and Oligo(dt)20 (ThermoFisher Scientific, 18418-020). .. RT-PCR and qPCR reactions were performed on Roche 480 (Roche, USA) using Power Up Sybr Green Master Mix (ThermoFisher; A25742) or TaqMan Universal PCR Mastermix for qPCR for CD16 (ThermoFisher Scientific, 4304437) for 40 cycles.

    Recombinant:

    Article Title: The vapB–vapC Operon of Acidovorax citrulli Functions as a Bona-fide Toxin–Antitoxin Module
    Article Snippet: In vitro Analyses of VapC RNase Activity 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).

    Isolation:

    Article Title: Circulating and Urinary miR-210 and miR-16 Increase during Cardiac Surgery Using Cardiopulmonary Bypass – A Pilot Study
    Article Snippet: RNA was extracted from urine (1 mL) using a Urine Exosome RNA Isolation Kit (Cat No 47200, Norgen Biotek Corp, Thorold, ON, Canada). .. After extraction of RNA, 10 μL of sample was pretreated with .5 μL of the RNase Inhibitor (Cat No M0307L, New England BioLabs® Inc, Ipswich, MA) and 2 units of Heparinase I (Cat No H2519, Sigma-Aldrich® , St Louis, MO) [in 20 mM Tris-HCl {pH 7.5}, 50 mM NaCl, 4 mM CaCl2 ] for 1 hour at 25°C to remove heparin ( ).

    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).

    Article Title: NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells
    Article Snippet: .. RT-PCR/qPCR RNA was isolated from canine expanded cells using RNAeasy Kit, QiaShredder Columns, and RNAase-Free DNase Set (all Qiagen, 74104, 79654, 79254), and cDNA was synthesized using Omniscript RT Kit (Qiagen, 20511) with RNase inhibitor (New England Biolabs, M0307S) and Oligo(dt)20 (ThermoFisher Scientific, 18418-020). .. RT-PCR and qPCR reactions were performed on Roche 480 (Roche, USA) using Power Up Sybr Green Master Mix (ThermoFisher; A25742) or TaqMan Universal PCR Mastermix for qPCR for CD16 (ThermoFisher Scientific, 4304437) for 40 cycles.

    Article Title: Lysine-specific Demethylase 2B (KDM2B)-let-7-Enhancer of Zester Homolog 2 (EZH2) Pathway Regulates Cell Cycle Progression and Senescence in Primary Cells *
    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′).

    Subcloning:

    Article Title: Detection of Endogenous MazF Enzymatic Activity in Staphylococcus aureus
    Article Snippet: Restriction enzymes, BSA, Low-Range ssRNA Ladder, RNase Inhibitor—Human Placenta, and E. coli strains DH5α and NiCo21(DE3) [ ] were purchased from New England Biolabs (NEB). .. Subcloning Efficiency DH5α Chemically Competent E. coli was purchased from Invitrogen.

    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 non-template control (RT-NTC) negative control reaction contained an equal volume of water instead of RNA.

    Purification:

    Article Title: Knockdown of fucosyltransferase III disrupts the adhesion of circulating cancer cells to E-selectin without affecting hematopoietic cell adhesion
    Article Snippet: Four days after siRNA transfection, total RNA was purified using RNeasy Plus mini Kit (Cat# 74134 Qiagen). .. The 40-μL reverse transcription reaction system included 5–10 μg of total RNA, 1 μL of M-MuLV Reverse Transcriptase (Cat# M0253L, New England Biolabs), 0.5 μL of RNase Inhibitor (Cat# M0307S, New England Biolabs), 1 μL of Random Primers (Cat# 48190-011, Invitrogen), 2.5 μL of dNTP Mix (Cat# 170-8874, New England Biolabs) and 4 μL of M-MuLV Reverse Transcriptase reaction buffer.

    Article Title: Data on the inhibition of RNase inhibitor activity by a monoclonal antibody as assessed by microfluidics-based RNA electrophoresis
    Article Snippet: .. 2.2 Inhibition of RI activity by a monoclonal Ab specific to RI (3F11) Purified total RNA from Vero cells (5 µg in 200 µL of CL Buffer) was mixed with 1 µg (1 µL) of monoclonal Ab specific to either RI (Origene; TA501875; clone 3F11) or GAPDH (Origene; TA802519; clone 2D9), 1 ng (1 µL) of RNase A (Qiagen; 19101; diluted in CL Buffer), 40 units (1 µL) of human placental RI (hpRI; New England Biolabs; M0307), or a combination of the above; in reactions containing hpRI, its addition preceded the addition of other components. .. Following the incubation, RNA was purified and subjected to Experion analysis.

    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: The expression and localization of RNase and RNase inhibitor in blood cells and vascular endothelial cells in homeostasis of the vascular system
    Article Snippet: .. Purified human RNase 1 (13468-H08H) and RI protein (M0307S) were from Sino Biologicals (Peking, PR China) and New England Biolabs Japan (Tokyo, Japan), respectively. .. Thrombin (Japan Blood Products Organization, Tokyo, Japan) was dissolved in normal saline and added to phosphate-buffered saline (PBS) to a final concentration of 2 nM.

    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

    Polymerase Chain Reaction:

    Article Title: Circulating and Urinary miR-210 and miR-16 Increase during Cardiac Surgery Using Cardiopulmonary Bypass – A Pilot Study
    Article Snippet: After extraction of RNA, 10 μL of sample was pretreated with .5 μL of the RNase Inhibitor (Cat No M0307L, New England BioLabs® Inc, Ipswich, MA) and 2 units of Heparinase I (Cat No H2519, Sigma-Aldrich® , St Louis, MO) [in 20 mM Tris-HCl {pH 7.5}, 50 mM NaCl, 4 mM CaCl2 ] for 1 hour at 25°C to remove heparin ( ). .. Preliminary experiments showed that heparinase does not affect polymerase chain reaction (PCR) analyses in serum and urine from non-heparinized patients, while allowing unaffected PCR amplification in samples potentially contaminated with heparin.

    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.

    Article Title: NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells
    Article Snippet: RT-PCR/qPCR RNA was isolated from canine expanded cells using RNAeasy Kit, QiaShredder Columns, and RNAase-Free DNase Set (all Qiagen, 74104, 79654, 79254), and cDNA was synthesized using Omniscript RT Kit (Qiagen, 20511) with RNase inhibitor (New England Biolabs, M0307S) and Oligo(dt)20 (ThermoFisher Scientific, 18418-020). .. RT-PCR and qPCR reactions were performed on Roche 480 (Roche, USA) using Power Up Sybr Green Master Mix (ThermoFisher; A25742) or TaqMan Universal PCR Mastermix for qPCR for CD16 (ThermoFisher Scientific, 4304437) for 40 cycles.

    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.

    Plasmid Preparation:

    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). .. The mixture was incubated for 2 hours at 37 °C to allow the in vitro transcription.

    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. .. 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) !

    In Vitro:

    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).

    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). .. The mixture was incubated for 2 hours at 37 °C to allow the in vitro transcription.

    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. .. 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) !

    SYBR Green Assay:

    Article Title: NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells
    Article Snippet: RT-PCR/qPCR RNA was isolated from canine expanded cells using RNAeasy Kit, QiaShredder Columns, and RNAase-Free DNase Set (all Qiagen, 74104, 79654, 79254), and cDNA was synthesized using Omniscript RT Kit (Qiagen, 20511) with RNase inhibitor (New England Biolabs, M0307S) and Oligo(dt)20 (ThermoFisher Scientific, 18418-020). .. RT-PCR and qPCR reactions were performed on Roche 480 (Roche, USA) using Power Up Sybr Green Master Mix (ThermoFisher; A25742) or TaqMan Universal PCR Mastermix for qPCR for CD16 (ThermoFisher Scientific, 4304437) for 40 cycles.

    RNA Extraction:

    Article Title: Circulating and Urinary miR-210 and miR-16 Increase during Cardiac Surgery Using Cardiopulmonary Bypass – A Pilot Study
    Article Snippet: Paragraph title: RNA Extraction ... After extraction of RNA, 10 μL of sample was pretreated with .5 μL of the RNase Inhibitor (Cat No M0307L, New England BioLabs® Inc, Ipswich, MA) and 2 units of Heparinase I (Cat No H2519, Sigma-Aldrich® , St Louis, MO) [in 20 mM Tris-HCl {pH 7.5}, 50 mM NaCl, 4 mM CaCl2 ] for 1 hour at 25°C to remove heparin ( ).

    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).

    Agarose Gel Electrophoresis:

    Article Title: NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells
    Article Snippet: RT-PCR/qPCR RNA was isolated from canine expanded cells using RNAeasy Kit, QiaShredder Columns, and RNAase-Free DNase Set (all Qiagen, 74104, 79654, 79254), and cDNA was synthesized using Omniscript RT Kit (Qiagen, 20511) with RNase inhibitor (New England Biolabs, M0307S) and Oligo(dt)20 (ThermoFisher Scientific, 18418-020). .. Semi-quantitative RT-PCR reactions were run on five different donors with 100 ng cDNA per sample and 6× DNA loading dye, and products were verified on a 1.5% agarose gel with TAE buffer and GelRed Nucleic Acid Stain (Phenix, RGB-4103) and 100 bp DNA Ladder (New England Biolabs, N3238S).

    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).

    Quantitation Assay:

    Article Title: Lysine-specific Demethylase 2B (KDM2B)-let-7-Enhancer of Zester Homolog 2 (EZH2) Pathway Regulates Cell Cycle Progression and Senescence in Primary Cells *
    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′).

    Concentration Assay:

    Article Title: The expression and localization of RNase and RNase inhibitor in blood cells and vascular endothelial cells in homeostasis of the vascular system
    Article Snippet: Purified human RNase 1 (13468-H08H) and RI protein (M0307S) were from Sino Biologicals (Peking, PR China) and New England Biolabs Japan (Tokyo, Japan), respectively. .. Thrombin (Japan Blood Products Organization, Tokyo, Japan) was dissolved in normal saline and added to phosphate-buffered saline (PBS) to a final concentration of 2 nM.

    Fractionation:

    Article Title: MitoRibo-Tag Mice Provide a Tool for In Vivo Studies of Mitoribosome Composition
    Article Snippet: Mitoribosome Gradient Analysis Sucrose density gradient fractionation of mitochondrial ribosomes was performed according to and . .. Mitochondria were resuspended at protein concentrations of 7.5 mg/ml (from cultured cells) or 10 mg/ml (from mouse tissues) in lysis buffer (260 mM sucrose, 10 mM Tris-HCl pH 7.5, 100 mM KCl, 20 mM MgCl2 , 2% digitonin for mouse mitochondria or 6% digitonin for human mitochondria, 40 U/ml RNase inhibitor (New England Biolabs, cat. no. M0307L) and 1x PIC) and incubated for 20 min on ice.

    Lysis:

    Article Title: MitoRibo-Tag Mice Provide a Tool for In Vivo Studies of Mitoribosome Composition
    Article Snippet: .. Mitochondria were resuspended at protein concentrations of 7.5 mg/ml (from cultured cells) or 10 mg/ml (from mouse tissues) in lysis buffer (260 mM sucrose, 10 mM Tris-HCl pH 7.5, 100 mM KCl, 20 mM MgCl2 , 2% digitonin for mouse mitochondria or 6% digitonin for human mitochondria, 40 U/ml RNase inhibitor (New England Biolabs, cat. no. M0307L) and 1x PIC) and incubated for 20 min on ice. .. Samples were clarified by centrifugation for 45 min/9200 g/4°C.

    Staining:

    Article Title: NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells
    Article Snippet: RT-PCR/qPCR RNA was isolated from canine expanded cells using RNAeasy Kit, QiaShredder Columns, and RNAase-Free DNase Set (all Qiagen, 74104, 79654, 79254), and cDNA was synthesized using Omniscript RT Kit (Qiagen, 20511) with RNase inhibitor (New England Biolabs, M0307S) and Oligo(dt)20 (ThermoFisher Scientific, 18418-020). .. Semi-quantitative RT-PCR reactions were run on five different donors with 100 ng cDNA per sample and 6× DNA loading dye, and products were verified on a 1.5% agarose gel with TAE buffer and GelRed Nucleic Acid Stain (Phenix, RGB-4103) and 100 bp DNA Ladder (New England Biolabs, N3238S).

    Hood:

    Article Title: Quantitative measurement of transcriptional inhibition and mutagenesis induced by site-specifically incorporated DNA lesions in vitro and in vivo
    Article Snippet: CAUTION Phenol:chloroform:isoamyl alcohol is toxic and corrosive; wear gloves and work in a fume hood. .. 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 rnase inhibitor
    Rnase Inhibitor, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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