sigma r6750  (Millipore)


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  • 99
    Name:
    Ribonucleic acid from baker s yeast
    Description:

    Catalog Number:
    r6750
    Price:
    None
    Applications:
    Ribonucleic acid (RNA) from baker's yeast S. cerevisiae may be used as a substrate for studying ribonuclease activities of enzymes such as RNAase(s).
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    Structured Review

    Millipore sigma r6750

    https://www.bioz.com/result/sigma r6750/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sigma r6750 - by Bioz Stars, 2020-04
    99/100 stars

    Related Products / Commonly Used Together

    yeast rna
    rnase a
    trna
    rrna
    vegf
    mrna
    ribonucleic acid

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    Related Articles

    Clone Assay:

    Article Title: Genetic ablation of butyrate utilization attenuates gastrointestinal Salmonella disease
    Article Snippet: .. Absolute values were calculated using a plasmid carrying the cloned gene to generate a standard curve ranging from 108 to 101 copies/ml diluted in a 0.02 mg/ml yeast RNA (Sigma-Aldrich, cat. # R6750) solution. ..

    Centrifugation:

    Article Title: Production and characterization of yeasts grown on media composed of spruce-derived sugars and protein hydrolysates from chicken by-products
    Article Snippet: Another 1.5 ml 0.5 M HClO4 was the added to the cells, followed by mixing, another incubation at 70 °C for 15 min, and centrifugation. .. HClO4 -extracts from the yeast cells and RNA standards (RNA from baker’s yeast; Sigma R6750) were mixed with a H2 SO4 /H2 O solution (v/v; 85/15) in 96 well plates, and incubated at 40 °C for 24 h. The orcinol reagent (stock solution of 0.35 mL 6% (w/v) orcinol to 5 mL concentrated HCl) was then added, and the plates were incubated with gentle shaking at 100 °C for 30 min, after which sbsorbance at 500 nm was read on a Spectramax M2e microplate reader (Molecular Devices).

    Blocking Assay:

    Article Title: Efficient detection of RNA-protein interactions using tethered RNAs
    Article Snippet: .. Egg white avidin (EMD chemicals, Cat#:189725, 10 µg/mg protein) and Yeast RNA (Sigma, Cat#R6750, ∼0.5 mg/mg protein) were added to block endogenous biotinylated proteins and non-specific RNPs, and incubated on a rotation shaker at 4°C, for 20 min. After blocking, the lysates were again centrifuged for 10 min at 16 000 g , 4°C, to prepare the final pre-cleared lysates. .. Supernatants were transferred to new tubes with 200 U/ml RNasin (Promega, cat#: N2115).

    Article Title: Sexually Dimorphic MicroRNA Expression During Chicken Embryonic Gonadal Development 1
    Article Snippet: .. Tissues were rehydrated through reverse MeOH/PBTX wash series (20 min per wash), washed two times for 10 min each with PBTX, then treated sequentially with proteinase K in PBTX (10 μg/ml) at room temperature for 30 min (E6.5) or 45 min (E9.5), received two 5-min PBTX washes, and then were refixed in 0.2% glutaraldehyde in 4% PFA/PBS for 20 min. Tissues were prehybridized at probe hybridization temperatures (22°C below probe Tm) for 2 h in 1.5–2 ml of hybridization solution (50% formamide; 5× saline-sodium citrate [SSC]; 2% blocking powder (Boehringer); 0.1% Triton X-100; 0.5% 3-[(cholamidopropyl)dimethylammonio]-1-propanesulfonate [CHAPS; Boehringer]; 1 ml of yeast RNA [no. R6750; Sigma]; 5 mM EDTA; and 50 μg/ml heparin). .. Prehybridized tissues were either hybridized immediately or stored at −20°C for 24–72 h. Prior to hybridization, stored prehybridized tissues were warmed to hybridization temperature for 2–4 h. DIG-labeled LNA probes were diluted to 2.5 pmol/μl and added to tissues at 2.5 μl per 1 ml of hybridization solution (typically 5 μl per tube of tissues).

    Adsorption:

    Article Title: Adsorption-Induced Changes in Ribonuclease A Structure and Enzymatic Activity on Solid Surfaces
    Article Snippet: .. Briefly, ribonucleic acid, which is the substrate for RNase A, was prepared in PPB to a final concentration of 20 mg/mL (Baker’s yeast, Sigma R6750) and exposed to RNase A both in solution and following RNase A adsorption. .. An initial calibration plot for solution-state enzymatic activity was obtained for a working range of 0.1–30 μg of RNAse A (based on the equivalently adsorbed amount of protein on different surfaces) by monitoring the absorbance at 300 nm (ΔA 300 ) at pH 7.4.

    Electrophoresis:

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: Paragraph title: 4.4. Electrophoresis Mobility Shift Assay ... RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750).

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: .. Electrophoresis Mobility Shift Assay RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Each sample, except for the total tRNA, was fragmented by sonication for better resolution in the electrophoresis mobility shift assay (EMSA).

    Quantitation Assay:

    Article Title: Impact of Soil Warming on the Plant Metabolome of Icelandic Grasslands
    Article Snippet: .. Reagents used were RNA (baker’s yeast S. cerevisiae , Sigma-Aldrich, Saint Louis, MO, USA) and DNA (calf thymus, Sigma-Aldrich, Saint Louis, MO, USA), RNase, DNase-free (Sigma-Aldrich, Saint Louis, MO, USA), N -lauroylsarcosine sodium salt (Sigma-Aldrich, Saint Louis, MO, USA), UltraPure™ DNase/RNase-Free Distilled Water (Life Technologies, Carlsbad, CA, USA), RiboGreen™ RNA Quantitation Kit (Molecular Probes, Inc., Eugene, OR, USA), PicoGreen™ dsDNA Quantitation Reagent (Molecular Probes, Inc., Eugene, OR, USA), TE buffer (RNase-free from RiboGreen™ RNA Quantitation Kit, Molecular Probes, Inc., Eugene, OR, USA), extraction buffer (1% w /v N -lauroylsarcosine sodium salt in TE buffer), and standard buffer (0.2% N -lauroylsarcosine sodium salt in TE buffer). ..

    Activity Assay:

    Article Title: Adsorption-Induced Changes in Ribonuclease A Structure and Enzymatic Activity on Solid Surfaces
    Article Snippet: Paragraph title: Characterization of Enzymatic Activity ... Briefly, ribonucleic acid, which is the substrate for RNase A, was prepared in PPB to a final concentration of 20 mg/mL (Baker’s yeast, Sigma R6750) and exposed to RNase A both in solution and following RNase A adsorption.

    Expressing:

    Article Title: Genetic ablation of butyrate utilization attenuates gastrointestinal Salmonella disease
    Article Snippet: Fold change in mRNA levels was determined using the comparative threshold cycle ( CT ) method and conditions were compared to either S. Typhimurium or S. Typhi in the absence of SCFAs for bacterial gene expression ( invA, hilD ) or mock-infected cecal tissue for murine gene expression experiments ( Il17a, Kc ). .. Absolute values were calculated using a plasmid carrying the cloned gene to generate a standard curve ranging from 108 to 101 copies/ml diluted in a 0.02 mg/ml yeast RNA (Sigma-Aldrich, cat. # R6750) solution.

    Modification:

    Article Title: Development of a Method to Extract Opium Poppy (Papaver somniferum L.) DNA from Heroin
    Article Snippet: Transfer Ribonucleic Acid (tRNA) from baker’s yeast ( S. cerevisiae ) and absolute ethanol were purchased from Sigma Chemical Company (St. Louis, MO). .. Several DNA extraction methods were evaluated during the course of this study: (1) the Qiagen Plant DNeasy Mini kit, (2) the Promega Maxwell 16 Tissue RNA LEV kit and (3) a modified version of the Qiagen Stool kit DNA extraction method.

    Hybridization:

    Article Title: Evidence that multiple P2X purinoceptors are functionally expressed in rat supraoptic neurones
    Article Snippet: .. Hybridization was carried out at 45°C overnight in 46 μl buffer (consisting of 42 μl of: 950 μl 1 M TrisCl pH 7.4, 190 μl 250 mM EDTA pH 8, 3.57 ml 4 M NaCl, 23.8 ml formamide, 9.53 ml 50 % dextran sulphate (500 000 MW), 950 μl Denhardt's solution and 1 ml water; with 2 μl of a solution of: 250 μl 10 mg ml−1 sheared salmon sperm DNA (D7656, Sigma), 250 μl 20 mg ml−1 yeast total RNA (type XI, R6750, Sigma) and 500 μl water; and ∼2 μl of 35 S-labelled antisense and sense probes (1 × 106 c.p.m. slide−1 )) under Nescofilm (Bando Chemical IMD, Ltd, Osaka, Japan) coverslips. .. The slides were rinsed in 2 × SSC (1 × SSC: 150 mM NaCl, 15 mM sodium citrate) and washed for 30 min in two changes of 2 × SSC-50 % formamide at 45°C before being incubated at 37°C in 2 × SSC containing 20 μg ml−1 RNase A (Sigma).

    Article Title: Sexually Dimorphic MicroRNA Expression During Chicken Embryonic Gonadal Development 1
    Article Snippet: .. Tissues were rehydrated through reverse MeOH/PBTX wash series (20 min per wash), washed two times for 10 min each with PBTX, then treated sequentially with proteinase K in PBTX (10 μg/ml) at room temperature for 30 min (E6.5) or 45 min (E9.5), received two 5-min PBTX washes, and then were refixed in 0.2% glutaraldehyde in 4% PFA/PBS for 20 min. Tissues were prehybridized at probe hybridization temperatures (22°C below probe Tm) for 2 h in 1.5–2 ml of hybridization solution (50% formamide; 5× saline-sodium citrate [SSC]; 2% blocking powder (Boehringer); 0.1% Triton X-100; 0.5% 3-[(cholamidopropyl)dimethylammonio]-1-propanesulfonate [CHAPS; Boehringer]; 1 ml of yeast RNA [no. R6750; Sigma]; 5 mM EDTA; and 50 μg/ml heparin). .. Prehybridized tissues were either hybridized immediately or stored at −20°C for 24–72 h. Prior to hybridization, stored prehybridized tissues were warmed to hybridization temperature for 2–4 h. DIG-labeled LNA probes were diluted to 2.5 pmol/μl and added to tissues at 2.5 μl per 1 ml of hybridization solution (typically 5 μl per tube of tissues).

    Mobility Shift:

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: Paragraph title: 4.4. Electrophoresis Mobility Shift Assay ... RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750).

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: .. Electrophoresis Mobility Shift Assay RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Each sample, except for the total tRNA, was fragmented by sonication for better resolution in the electrophoresis mobility shift assay (EMSA).

    Transferring:

    Article Title: Evidence that multiple P2X purinoceptors are functionally expressed in rat supraoptic neurones
    Article Snippet: Fixed tissue sections were then incubated for 10 min in 0.9 % saline containing 0.1 M triethanolamine and 0.25 % acetic anhydride, dehydrated by transferring them sequentially through 70 % (1 min), 80 % (1 min), 95 % (2 min) and 100 % ethanol (1 min), and delipidated in 100 % chloroform for 5 min. .. Hybridization was carried out at 45°C overnight in 46 μl buffer (consisting of 42 μl of: 950 μl 1 M TrisCl pH 7.4, 190 μl 250 mM EDTA pH 8, 3.57 ml 4 M NaCl, 23.8 ml formamide, 9.53 ml 50 % dextran sulphate (500 000 MW), 950 μl Denhardt's solution and 1 ml water; with 2 μl of a solution of: 250 μl 10 mg ml−1 sheared salmon sperm DNA (D7656, Sigma), 250 μl 20 mg ml−1 yeast total RNA (type XI, R6750, Sigma) and 500 μl water; and ∼2 μl of 35 S-labelled antisense and sense probes (1 × 106 c.p.m. slide−1 )) under Nescofilm (Bando Chemical IMD, Ltd, Osaka, Japan) coverslips.

    Generated:

    Article Title: Toward a General Approach for RNA-Templated Hierarchical Assembly of Split-Proteins
    Article Snippet: CLuciferase-E2C and Aart-NLuciferase mRNA transcripts were generated as described above. .. For detection of VEGF in the presence of yeast RNA (Ribonucleic acid mixture from baker's yeast ( S. cerevisiae ) containing mRNA, tRNA, and rRNA, Sigma R6750), translations were performed followed by addition of 500 pM (5.0 ng) annealed VEGF RNA, 500 pM annealed VEGF + 2000-fold mass excess (10 μg) yeast RNA, yeast RNA + HP-guides, or HP-guides only.

    Polymerase Chain Reaction:

    Article Title: Development of a Method to Extract Opium Poppy (Papaver somniferum L.) DNA from Heroin
    Article Snippet: Fast Start High Fidelity PCR System, dNTP packs were purchased from Roche Life Science (Indianapolis, IN). .. Transfer Ribonucleic Acid (tRNA) from baker’s yeast ( S. cerevisiae ) and absolute ethanol were purchased from Sigma Chemical Company (St. Louis, MO).

    Article Title: Sexually Dimorphic MicroRNA Expression During Chicken Embryonic Gonadal Development 1
    Article Snippet: The UGSs were dissected from E6.5 and E9.5 embryos, sexed by PCR, and fixed overnight in 4% paraformaldehyde (PFA) in DEPC-PBS. .. Tissues were rehydrated through reverse MeOH/PBTX wash series (20 min per wash), washed two times for 10 min each with PBTX, then treated sequentially with proteinase K in PBTX (10 μg/ml) at room temperature for 30 min (E6.5) or 45 min (E9.5), received two 5-min PBTX washes, and then were refixed in 0.2% glutaraldehyde in 4% PFA/PBS for 20 min. Tissues were prehybridized at probe hybridization temperatures (22°C below probe Tm) for 2 h in 1.5–2 ml of hybridization solution (50% formamide; 5× saline-sodium citrate [SSC]; 2% blocking powder (Boehringer); 0.1% Triton X-100; 0.5% 3-[(cholamidopropyl)dimethylammonio]-1-propanesulfonate [CHAPS; Boehringer]; 1 ml of yeast RNA [no. R6750; Sigma]; 5 mM EDTA; and 50 μg/ml heparin).

    Article Title: Genetic ablation of butyrate utilization attenuates gastrointestinal Salmonella disease
    Article Snippet: PCR mix and the appropriate primer sets ( ) at a final concentration of 0.25 mM. .. Absolute values were calculated using a plasmid carrying the cloned gene to generate a standard curve ranging from 108 to 101 copies/ml diluted in a 0.02 mg/ml yeast RNA (Sigma-Aldrich, cat. # R6750) solution.

    Sonication:

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Each sample, except for the total tRNA, was fragmented by sonication for better resolution in the electrophoresis mobility shift assay (EMSA).

    Article Title: Efficient detection of RNA-protein interactions using tethered RNAs
    Article Snippet: Total cell lysates were lightly sonicated, and centrifuged for 10 min at 16 000g , 4°C. .. Egg white avidin (EMD chemicals, Cat#:189725, 10 µg/mg protein) and Yeast RNA (Sigma, Cat#R6750, ∼0.5 mg/mg protein) were added to block endogenous biotinylated proteins and non-specific RNPs, and incubated on a rotation shaker at 4°C, for 20 min. After blocking, the lysates were again centrifuged for 10 min at 16 000 g , 4°C, to prepare the final pre-cleared lysates.

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: Electrophoresis Mobility Shift Assay RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Each sample, except for the total tRNA, was fragmented by sonication for better resolution in the electrophoresis mobility shift assay (EMSA).

    Binding Assay:

    Article Title: Toward a General Approach for RNA-Templated Hierarchical Assembly of Split-Proteins
    Article Snippet: For detection of VEGF in the presence of yeast RNA (Ribonucleic acid mixture from baker's yeast ( S. cerevisiae ) containing mRNA, tRNA, and rRNA, Sigma R6750), translations were performed followed by addition of 500 pM (5.0 ng) annealed VEGF RNA, 500 pM annealed VEGF + 2000-fold mass excess (10 μg) yeast RNA, yeast RNA + HP-guides, or HP-guides only. .. However, to determine a time-dependence of biosensor binding to the ternary VEGF target, incubation times were 5, 10, 20, 30, 45, or 60 minutes.

    DNA Extraction:

    Article Title: Development of a Method to Extract Opium Poppy (Papaver somniferum L.) DNA from Heroin
    Article Snippet: Transfer Ribonucleic Acid (tRNA) from baker’s yeast ( S. cerevisiae ) and absolute ethanol were purchased from Sigma Chemical Company (St. Louis, MO). .. Several DNA extraction methods were evaluated during the course of this study: (1) the Qiagen Plant DNeasy Mini kit, (2) the Promega Maxwell 16 Tissue RNA LEV kit and (3) a modified version of the Qiagen Stool kit DNA extraction method.

    Article Title: Genetic ablation of butyrate utilization attenuates gastrointestinal Salmonella disease
    Article Snippet: DNA from the cecal contents was extracted using the PowerSoil DNA Isolation kit according to the manufacturer’s protocol. .. Absolute values were calculated using a plasmid carrying the cloned gene to generate a standard curve ranging from 108 to 101 copies/ml diluted in a 0.02 mg/ml yeast RNA (Sigma-Aldrich, cat. # R6750) solution.

    Pull Down Assay:

    Article Title: Efficient detection of RNA-protein interactions using tethered RNAs
    Article Snippet: Paragraph title: RNA pull-down assay ... Egg white avidin (EMD chemicals, Cat#:189725, 10 µg/mg protein) and Yeast RNA (Sigma, Cat#R6750, ∼0.5 mg/mg protein) were added to block endogenous biotinylated proteins and non-specific RNPs, and incubated on a rotation shaker at 4°C, for 20 min. After blocking, the lysates were again centrifuged for 10 min at 16 000 g , 4°C, to prepare the final pre-cleared lysates.

    Avidin-Biotin Assay:

    Article Title: Efficient detection of RNA-protein interactions using tethered RNAs
    Article Snippet: .. Egg white avidin (EMD chemicals, Cat#:189725, 10 µg/mg protein) and Yeast RNA (Sigma, Cat#R6750, ∼0.5 mg/mg protein) were added to block endogenous biotinylated proteins and non-specific RNPs, and incubated on a rotation shaker at 4°C, for 20 min. After blocking, the lysates were again centrifuged for 10 min at 16 000 g , 4°C, to prepare the final pre-cleared lysates. .. Supernatants were transferred to new tubes with 200 U/ml RNasin (Promega, cat#: N2115).

    Article Title: Detection of RNA-Protein Interactions using Tethered RNA Affinity Capture
    Article Snippet: .. White Egg Avidin (EMD chemicals, cat#189725) Yeast RNA (Sigma, cat#R6750) Streptavidin agarose (Pierce, cat#20349) ..

    Microscopy:

    Article Title: Evidence that multiple P2X purinoceptors are functionally expressed in rat supraoptic neurones
    Article Snippet: Hybridization was carried out at 45°C overnight in 46 μl buffer (consisting of 42 μl of: 950 μl 1 M TrisCl pH 7.4, 190 μl 250 mM EDTA pH 8, 3.57 ml 4 M NaCl, 23.8 ml formamide, 9.53 ml 50 % dextran sulphate (500 000 MW), 950 μl Denhardt's solution and 1 ml water; with 2 μl of a solution of: 250 μl 10 mg ml−1 sheared salmon sperm DNA (D7656, Sigma), 250 μl 20 mg ml−1 yeast total RNA (type XI, R6750, Sigma) and 500 μl water; and ∼2 μl of 35 S-labelled antisense and sense probes (1 × 106 c.p.m. slide−1 )) under Nescofilm (Bando Chemical IMD, Ltd, Osaka, Japan) coverslips. .. Slides hybridized to the riboprobes were dipped in nuclear emulsion (K-5, Ilford, Cheshire, UK) and exposed for 14 days before being developed in D-19 developer (Kodak, Rochester, NY, USA), fixed with Fujifix (Fuji Photo Film Co., Ltd, Tokyo, Japan) and examined with bright- and dark-field microscopy.

    Purification:

    Article Title: Development of a Method to Extract Opium Poppy (Papaver somniferum L.) DNA from Heroin
    Article Snippet: Maxwell 16 Tissue LEV RNA purification kits were purchased from Promega (Madison, WI). .. Transfer Ribonucleic Acid (tRNA) from baker’s yeast ( S. cerevisiae ) and absolute ethanol were purchased from Sigma Chemical Company (St. Louis, MO).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Efficient detection of RNA-protein interactions using tethered RNAs
    Article Snippet: .. Egg white avidin (EMD chemicals, Cat:189725, 10 µg/mg protein) and Yeast RNA (Sigma, Cat#R6750, ∼0.5 mg/mg protein) were added to block endogenous biotinylated proteins and non-specific RNPs, and incubated on a rotation shaker at 4°C, for 20 min. After blocking, the lysates were again centrifuged for 10 min at 16 000 g , 4°C, to prepare the final pre-cleared lysates. .. Supernatants were transferred to new tubes with 200 U/ml RNasin (Promega, cat#: N2115).

    Article Title: Detection of RNA-Protein Interactions using Tethered RNA Affinity Capture
    Article Snippet: .. White Egg Avidin (EMD chemicals, cat189725) Yeast RNA (Sigma, cat#R6750) Streptavidin agarose (Pierce, cat#20349) ..

    In Situ Hybridization:

    Article Title: Evidence that multiple P2X purinoceptors are functionally expressed in rat supraoptic neurones
    Article Snippet: Paragraph title: In situ hybridization histochemistry ... Hybridization was carried out at 45°C overnight in 46 μl buffer (consisting of 42 μl of: 950 μl 1 M TrisCl pH 7.4, 190 μl 250 mM EDTA pH 8, 3.57 ml 4 M NaCl, 23.8 ml formamide, 9.53 ml 50 % dextran sulphate (500 000 MW), 950 μl Denhardt's solution and 1 ml water; with 2 μl of a solution of: 250 μl 10 mg ml−1 sheared salmon sperm DNA (D7656, Sigma), 250 μl 20 mg ml−1 yeast total RNA (type XI, R6750, Sigma) and 500 μl water; and ∼2 μl of 35 S-labelled antisense and sense probes (1 × 106 c.p.m. slide−1 )) under Nescofilm (Bando Chemical IMD, Ltd, Osaka, Japan) coverslips.

    Plasmid Preparation:

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: .. RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Each sample, except for the total tRNA, was fragmented by sonication for better resolution in the electrophoresis mobility shift assay (EMSA).

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: .. Electrophoresis Mobility Shift Assay RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Each sample, except for the total tRNA, was fragmented by sonication for better resolution in the electrophoresis mobility shift assay (EMSA).

    Article Title: Genetic ablation of butyrate utilization attenuates gastrointestinal Salmonella disease
    Article Snippet: .. Absolute values were calculated using a plasmid carrying the cloned gene to generate a standard curve ranging from 108 to 101 copies/ml diluted in a 0.02 mg/ml yeast RNA (Sigma-Aldrich, cat. # R6750) solution. ..

    Spectrophotometric Assay:

    Article Title: Adsorption-Induced Changes in Ribonuclease A Structure and Enzymatic Activity on Solid Surfaces
    Article Snippet: II.e Characterization of Enzymatic Activity A spectrophotometric assay was used to measure the enzymatic activity of RNase A to complement the CD and AAL/MS data. .. Briefly, ribonucleic acid, which is the substrate for RNase A, was prepared in PPB to a final concentration of 20 mg/mL (Baker’s yeast, Sigma R6750) and exposed to RNase A both in solution and following RNase A adsorption.

    Real-time Polymerase Chain Reaction:

    Article Title: Genetic ablation of butyrate utilization attenuates gastrointestinal Salmonella disease
    Article Snippet: Paragraph title: Quantitative real-time PCR analysis. ... Absolute values were calculated using a plasmid carrying the cloned gene to generate a standard curve ranging from 108 to 101 copies/ml diluted in a 0.02 mg/ml yeast RNA (Sigma-Aldrich, cat. # R6750) solution.

    Agarose Gel Electrophoresis:

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Total tRNA and fragmented NAs were mixed with the defined amount of hRID, incubated for 10 min at room temperature, and analyzed by agarose gel electrophoresis.

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: Electrophoresis Mobility Shift Assay RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Total tRNA and fragmented NAs were mixed with the defined amount of hRID, incubated for 10 min at room temperature, and analyzed by agarose gel electrophoresis.

    Radial Immuno Diffusion:

    Article Title: Evidence that multiple P2X purinoceptors are functionally expressed in rat supraoptic neurones
    Article Snippet: .. Hybridization was carried out at 45°C overnight in 46 μl buffer (consisting of 42 μl of: 950 μl 1 M TrisCl pH 7.4, 190 μl 250 mM EDTA pH 8, 3.57 ml 4 M NaCl, 23.8 ml formamide, 9.53 ml 50 % dextran sulphate (500 000 MW), 950 μl Denhardt's solution and 1 ml water; with 2 μl of a solution of: 250 μl 10 mg ml−1 sheared salmon sperm DNA (D7656, Sigma), 250 μl 20 mg ml−1 yeast total RNA (type XI, R6750, Sigma) and 500 μl water; and ∼2 μl of 35 S-labelled antisense and sense probes (1 × 106 c.p.m. slide−1 )) under Nescofilm (Bando Chemical IMD, Ltd, Osaka, Japan) coverslips. .. The slides were rinsed in 2 × SSC (1 × SSC: 150 mM NaCl, 15 mM sodium citrate) and washed for 30 min in two changes of 2 × SSC-50 % formamide at 45°C before being incubated at 37°C in 2 × SSC containing 20 μg ml−1 RNase A (Sigma).

    Incubation:

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Total tRNA and fragmented NAs were mixed with the defined amount of hRID, incubated for 10 min at room temperature, and analyzed by agarose gel electrophoresis.

    Article Title: Efficient detection of RNA-protein interactions using tethered RNAs
    Article Snippet: .. Egg white avidin (EMD chemicals, Cat#:189725, 10 µg/mg protein) and Yeast RNA (Sigma, Cat#R6750, ∼0.5 mg/mg protein) were added to block endogenous biotinylated proteins and non-specific RNPs, and incubated on a rotation shaker at 4°C, for 20 min. After blocking, the lysates were again centrifuged for 10 min at 16 000 g , 4°C, to prepare the final pre-cleared lysates. .. Supernatants were transferred to new tubes with 200 U/ml RNasin (Promega, cat#: N2115).

    Article Title: Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase
    Article Snippet: Electrophoresis Mobility Shift Assay RNA-free hRID was mixed with various types of NAs including total tRNA from E. coli (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. R-1753), pGE backbone plasmid DNA, polyA + polyU mixture (Pharmacia Biotech, Township, NJ, USA; Cat. No. 27-4110-01 and 27-4440-02, respectively), calf thymus DNA (Sigma-Aldrich; 27-4563-01), and yeast RNA (Sigma-Aldrich; R-6750). .. Total tRNA and fragmented NAs were mixed with the defined amount of hRID, incubated for 10 min at room temperature, and analyzed by agarose gel electrophoresis.

    Article Title: Toward a General Approach for RNA-Templated Hierarchical Assembly of Split-Proteins
    Article Snippet: For detection of VEGF in the presence of yeast RNA (Ribonucleic acid mixture from baker's yeast ( S. cerevisiae ) containing mRNA, tRNA, and rRNA, Sigma R6750), translations were performed followed by addition of 500 pM (5.0 ng) annealed VEGF RNA, 500 pM annealed VEGF + 2000-fold mass excess (10 μg) yeast RNA, yeast RNA + HP-guides, or HP-guides only. .. In most cases luminescence readings were taken following 30 minutes of incubation with the ternary complex at 4 °C.

    Article Title: Production and characterization of yeasts grown on media composed of spruce-derived sugars and protein hydrolysates from chicken by-products
    Article Snippet: .. HClO4 -extracts from the yeast cells and RNA standards (RNA from baker’s yeast; Sigma R6750) were mixed with a H2 SO4 /H2 O solution (v/v; 85/15) in 96 well plates, and incubated at 40 °C for 24 h. The orcinol reagent (stock solution of 0.35 mL 6% (w/v) orcinol to 5 mL concentrated HCl) was then added, and the plates were incubated with gentle shaking at 100 °C for 30 min, after which sbsorbance at 500 nm was read on a Spectramax M2e microplate reader (Molecular Devices). ..

    Article Title: Evidence that multiple P2X purinoceptors are functionally expressed in rat supraoptic neurones
    Article Snippet: Fixed tissue sections were then incubated for 10 min in 0.9 % saline containing 0.1 M triethanolamine and 0.25 % acetic anhydride, dehydrated by transferring them sequentially through 70 % (1 min), 80 % (1 min), 95 % (2 min) and 100 % ethanol (1 min), and delipidated in 100 % chloroform for 5 min. .. Hybridization was carried out at 45°C overnight in 46 μl buffer (consisting of 42 μl of: 950 μl 1 M TrisCl pH 7.4, 190 μl 250 mM EDTA pH 8, 3.57 ml 4 M NaCl, 23.8 ml formamide, 9.53 ml 50 % dextran sulphate (500 000 MW), 950 μl Denhardt's solution and 1 ml water; with 2 μl of a solution of: 250 μl 10 mg ml−1 sheared salmon sperm DNA (D7656, Sigma), 250 μl 20 mg ml−1 yeast total RNA (type XI, R6750, Sigma) and 500 μl water; and ∼2 μl of 35 S-labelled antisense and sense probes (1 × 106 c.p.m. slide−1 )) under Nescofilm (Bando Chemical IMD, Ltd, Osaka, Japan) coverslips.

    Produced:

    Article Title: Genetic ablation of butyrate utilization attenuates gastrointestinal Salmonella disease
    Article Snippet: As a control, bacterial target gene expression was normalized to the S. Typhimurium rpoA gene message, which produced similar results as normalization with the 16S rRNA gene message (data not shown). .. Absolute values were calculated using a plasmid carrying the cloned gene to generate a standard curve ranging from 108 to 101 copies/ml diluted in a 0.02 mg/ml yeast RNA (Sigma-Aldrich, cat. # R6750) solution.

    Concentration Assay:

    Article Title: Adsorption-Induced Changes in Ribonuclease A Structure and Enzymatic Activity on Solid Surfaces
    Article Snippet: .. Briefly, ribonucleic acid, which is the substrate for RNase A, was prepared in PPB to a final concentration of 20 mg/mL (Baker’s yeast, Sigma R6750) and exposed to RNase A both in solution and following RNase A adsorption. .. An initial calibration plot for solution-state enzymatic activity was obtained for a working range of 0.1–30 μg of RNAse A (based on the equivalently adsorbed amount of protein on different surfaces) by monitoring the absorbance at 300 nm (ΔA 300 ) at pH 7.4.

    Article Title: Genetic ablation of butyrate utilization attenuates gastrointestinal Salmonella disease
    Article Snippet: PCR mix and the appropriate primer sets ( ) at a final concentration of 0.25 mM. .. Absolute values were calculated using a plasmid carrying the cloned gene to generate a standard curve ranging from 108 to 101 copies/ml diluted in a 0.02 mg/ml yeast RNA (Sigma-Aldrich, cat. # R6750) solution.

    Lysis:

    Article Title: Efficient detection of RNA-protein interactions using tethered RNAs
    Article Snippet: Lysis buffer was prepared with 0.1% DEP-treated water. .. Egg white avidin (EMD chemicals, Cat#:189725, 10 µg/mg protein) and Yeast RNA (Sigma, Cat#R6750, ∼0.5 mg/mg protein) were added to block endogenous biotinylated proteins and non-specific RNPs, and incubated on a rotation shaker at 4°C, for 20 min. After blocking, the lysates were again centrifuged for 10 min at 16 000 g , 4°C, to prepare the final pre-cleared lysates.

    Article Title: Detection of RNA-Protein Interactions using Tethered RNA Affinity Capture
    Article Snippet: Lysis Buffer: 10 mM HEPES pH 7.0, 200 mM NaCl, 10 mM MgCl2 , 1 % TritonX-100, 1 mM DTT (Dithiothreitol), and protease inhibitors. .. White Egg Avidin (EMD chemicals, cat#189725) Yeast RNA (Sigma, cat#R6750) Streptavidin agarose (Pierce, cat#20349)