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Promega recombinant rnasin rnase inhibitor
Recombinant Rnasin Rnase Inhibitor, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 38 article reviews
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recombinant rnasin rnase inhibitor - by Bioz Stars, 2020-03
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Centrifugation:

Article Title: Codon-usage-based inhibition of HIV protein synthesis by human schlafen 11
Article Snippet: Briefly, freshly harvested cells were gently lysed on ice for 5 min in buffer RLN (50 mM Tris·Cl, pH 8.0, 140 mM NaCl, 1.5 mM MgCl2 , 0.5% (v/v) IGEPAL CA-630, 1,000 U ml−1 RNasin Plus RNase Inhibitor (Promega) and 1 mM DTT). .. Intact cell nuclei were removed by centrifugation at 4 °C for 2 min at 300 g , and cytoplasmic RNA was extracted from the remaining supernatant.

Amplification:

Article Title: Telomerase in kinetoplastid parasitic protozoa
Article Snippet: Paragraph title: Telomere Repeat Amplification Protocol (TRAP). ... Activity in the extracts was tested for RNase A sensitivity by incubation with 100 ng of RNase A (Sigma) for 5 min at 37°C before or after the telomerase reaction step and with or without 1 unit of RNase inhibitor RNasin (Promega) before addition of RNase A.

Article Title: Chicken Cells Sense Influenza A Virus Infection through MDA5 and CARDIF Signaling Involving LGP2
Article Snippet: .. RNA was extracted using TRIzol (Invitrogen), DNase I (Ambion), and RNase inhibitor (RNasin Plus; Promega) or with the Nucleospin RNA II kit (Macherey-Nagel) and amplified with the SuperScript III platinum one-step quantitative RT-PCR system (Invitrogen) using oligonucleotide primers and probes ( ) purchased from Microsynth. ..

Quantitative RT-PCR:

Article Title: Chicken Cells Sense Influenza A Virus Infection through MDA5 and CARDIF Signaling Involving LGP2
Article Snippet: .. RNA was extracted using TRIzol (Invitrogen), DNase I (Ambion), and RNase inhibitor (RNasin Plus; Promega) or with the Nucleospin RNA II kit (Macherey-Nagel) and amplified with the SuperScript III platinum one-step quantitative RT-PCR system (Invitrogen) using oligonucleotide primers and probes ( ) purchased from Microsynth. ..

Real-time Polymerase Chain Reaction:

Article Title: Codon-usage-based inhibition of HIV protein synthesis by human schlafen 11
Article Snippet: Paragraph title: Quantitative PCR and primers ... Briefly, freshly harvested cells were gently lysed on ice for 5 min in buffer RLN (50 mM Tris·Cl, pH 8.0, 140 mM NaCl, 1.5 mM MgCl2 , 0.5% (v/v) IGEPAL CA-630, 1,000 U ml−1 RNasin Plus RNase Inhibitor (Promega) and 1 mM DTT).

Incubation:

Article Title: ALS mutant FUS disrupts nuclear localization and sequesters wild-type FUS within cytoplasmic stress granules
Article Snippet: .. 5 µl of an RNase A/T1 mix (Fermentas, Yorkshire, UK) or RNasin Plus RNase inhibitor (Promega, Southampton, UK) was added to the tubes and incubated at 37°C for 5 min, then placed back on ice. .. Mouse anti-Myc (Cell Signalling Technology, 1:100), rabbit anti-HA (Cell Signalling Technology, 1:100) or mouse anti-FUS (Santa Cruz Biotechnology, Inc., 1:100) was added to the lysates as appropriate followed by the addition of 20 µl of protein A beads (Invitrogen Dynabeads).

Article Title: The RNA Capping Enzyme Domain in Protein A is Essential for Flock House Virus Replication
Article Snippet: In vitro transcribed uncapped T_Rluc and FHV_T RNA were made by assembling on ice 750 ng linearized DNA, a final concentration of 1 mM (each) ATP/CTP/GTP/UTP, 5 mM dithiothreitol (DTT), transcription buffer (Promega), 0.2 U/µL RNasin ribonuclease inhibitor (Promega) and 40 U T7 RNA polymerase (Promega) in a final volume of 50 µL. .. The reaction was incubated for 2 h at 37 °C, after which 2 µL of RNase-free DNaseI (Promega) was added and further incubated for 35 min.

Article Title: The structure of human Nocturnin reveals a conserved ribonuclease domain that represses target transcript translation and abundance in cells
Article Snippet: .. NOCT64–431 , CNOT6L158–555 or CNOT6L158–555 E240A (1 μM) were mixed with 10 nM 5′ Cy3 labeled p(A)20 (Dharmacon), and RNasin RNase A inhibitor (Promega) and incubated over a 30-min time course at 37°C. ..

Article Title: Increased Hematopoietic Extracellular RNAs and Vesicles in the Lung during Allergic Airway Responses
Article Snippet: .. exRNA stability To test whether synthetic miRNA-like oligoribonucleotide sequences were intrinsically stable in BALF, 22 nucleotide RNA calibrators (calibrators 1-4 ( )) were spiked into BALF and incubated for 5min at 37°C with or without RNAsin ribonuclease inhibitor (Promega) before RNA extraction. .. To test whether endogenous ex-miRNAs were stable to exogenous RNases, RNase A (Thermo Scientific) was added to BALF or total RNA previously isolated from BALF at 0.04-40ug/ml and incubated for 37°C for 15min before RNA extraction.

Article Title: Role for Bovine Viral Diarrhea Virus Erns Glycoprotein in the Control of Activation of Beta Interferon by Double-Stranded RNA
Article Snippet: The glycoproteins in cell culture medium were further incubated at 37°C for 18 h, and the RNase activity was determined as described previously ( ) with slight modification. .. Briefly, the assay mixture (25 μl in total) contained 0.2 μg of wild-type or mutant Erns , 12.5 μg of yeast 16-23S RNA (Worthington), and 40 U of RNase inhibitor (RNasin; Promega) in 20 mM sodium acetate buffer (pH 4.5).

Article Title: Telomerase in kinetoplastid parasitic protozoa
Article Snippet: .. Activity in the extracts was tested for RNase A sensitivity by incubation with 100 ng of RNase A (Sigma) for 5 min at 37°C before or after the telomerase reaction step and with or without 1 unit of RNase inhibitor RNasin (Promega) before addition of RNase A. ..

Article Title: Increased Hematopoietic Extracellular RNAs and Vesicles in the Lung during Allergic Airway Responses
Article Snippet: .. To test whether synthetic miRNA-like oligoribonucleotide sequences were intrinsically stable in BALF, 22 nucleotide RNA calibrators (calibrators 1-4 ( )) were spiked into BALF and incubated for 5min at 37°C with or without RNAsin ribonuclease inhibitor (Promega) before RNA extraction. .. To test whether endogenous ex-miRNAs were stable to exogenous RNases, RNase A (Thermo Scientific) was added to BALF or total RNA previously isolated from BALF at 0.04-40ug/ml and incubated for 37°C for 15min before RNA extraction.

Gel Extraction:

Article Title: The RNA Capping Enzyme Domain in Protein A is Essential for Flock House Virus Replication
Article Snippet: The DNA band was excised from the gel and extracted using GeneJET gel extraction kit (Thermo Fisher Scientific) as per the manufacturer’s instructions. .. In vitro transcribed uncapped T_Rluc and FHV_T RNA were made by assembling on ice 750 ng linearized DNA, a final concentration of 1 mM (each) ATP/CTP/GTP/UTP, 5 mM dithiothreitol (DTT), transcription buffer (Promega), 0.2 U/µL RNasin ribonuclease inhibitor (Promega) and 40 U T7 RNA polymerase (Promega) in a final volume of 50 µL.

Activity Assay:

Article Title: Role for Bovine Viral Diarrhea Virus Erns Glycoprotein in the Control of Activation of Beta Interferon by Double-Stranded RNA
Article Snippet: Paragraph title: RNase activity assay. ... Briefly, the assay mixture (25 μl in total) contained 0.2 μg of wild-type or mutant Erns , 12.5 μg of yeast 16-23S RNA (Worthington), and 40 U of RNase inhibitor (RNasin; Promega) in 20 mM sodium acetate buffer (pH 4.5).

Article Title: Telomerase in kinetoplastid parasitic protozoa
Article Snippet: .. Activity in the extracts was tested for RNase A sensitivity by incubation with 100 ng of RNase A (Sigma) for 5 min at 37°C before or after the telomerase reaction step and with or without 1 unit of RNase inhibitor RNasin (Promega) before addition of RNase A. ..

Article Title: Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
Article Snippet: .. RNase P activity assay RNase P cleavage was performed by mixing 300 nm MRPP1–MRPP2, 150 nm MRPP3, 10 units of RNase inhibitors (RNasin from Promega), and 400 nm in vitro transcribed pre-(mt) tRNAIle in a buffer of 30 mm Tris-HCl, pH 8, 40 mm NaCl, 4.5 mm MgCl2 , and 2 mm DTT to a total reaction volume of 8.25 μl. .. The reaction was performed at room temperature and stopped at set times by transferring 1 μl of the reaction mixture into 5 μl of 500 mm EDTA and heating to 95 °C.

Modification:

Article Title: Role for Bovine Viral Diarrhea Virus Erns Glycoprotein in the Control of Activation of Beta Interferon by Double-Stranded RNA
Article Snippet: The glycoproteins in cell culture medium were further incubated at 37°C for 18 h, and the RNase activity was determined as described previously ( ) with slight modification. .. Briefly, the assay mixture (25 μl in total) contained 0.2 μg of wild-type or mutant Erns , 12.5 μg of yeast 16-23S RNA (Worthington), and 40 U of RNase inhibitor (RNasin; Promega) in 20 mM sodium acetate buffer (pH 4.5).

Transfection:

Article Title: ALS mutant FUS disrupts nuclear localization and sequesters wild-type FUS within cytoplasmic stress granules
Article Snippet: Co-immunoprecipitation HEK 293T cells were grown in 10 cm dishes and then transfected using FuGene HD (Roche) according to the manufacturer's protocol. .. 5 µl of an RNase A/T1 mix (Fermentas, Yorkshire, UK) or RNasin Plus RNase inhibitor (Promega, Southampton, UK) was added to the tubes and incubated at 37°C for 5 min, then placed back on ice.

Ligation:

Article Title: Selective amplification and sequencing of cyclic phosphate-containing RNAs by the cP-RNA-seq method
Article Snippet: Sodium Acetate (3 M), pH 5.5 (Life Technologies, cat. no. AM9740) EDTA (0.5 M), pH 8.0 (Life Technologies, cat. no. AM9260G) SDS, 20% Solution (Life Technologies, cat. no. AM9820) Alkaline Phosphatase, Calf Intestinal (CIP; New England Biolabs Inc, cat. no. M0290) 10× CutSmart Buffer (supplied with CIP) RNasin® Ribonuclease Inhibitor (Promega, cat. no. N2111) Acid-Phenol:Chloroform, pH 4.5 (with IAA, 125:24:1) (Life Technologies, cat. no. AM9720) CAUTION! .. Linear Acrylamide (Life Technologies, cat. no. AM9520) Sodium periodate (Sigma-Aldrich, cat. no. 311448) T4 Polynucleotide Kinase (T4 PNK; New England Biolabs Inc, cat. no. M0201) 10× T4 Polynucleotide Kinase Reaction Buffer (supplied with T4 PNK) Adenosine 5′-Triphosphate (ATP) (New England Biolabs Inc, cat. no. P0756) TruSeq Small RNA Library Preparation Kits (Illumina, cat. no. RS-200-0012), containing RNA 3′ Adapter, Ligation Buffer, Stop Solution, RNA 5′ Adapter, 10 mM ATP, T4 RNA Ligase, 25 mM dNTP mix, RNA RT Primer, PCR Mix, RNA PCR Primer, RNA PCR Primer Index, and High Resolution Ladder.

Transferring:

Article Title: Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
Article Snippet: RNase P activity assay RNase P cleavage was performed by mixing 300 nm MRPP1–MRPP2, 150 nm MRPP3, 10 units of RNase inhibitors (RNasin from Promega), and 400 nm in vitro transcribed pre-(mt) tRNAIle in a buffer of 30 mm Tris-HCl, pH 8, 40 mm NaCl, 4.5 mm MgCl2 , and 2 mm DTT to a total reaction volume of 8.25 μl. .. The reaction was performed at room temperature and stopped at set times by transferring 1 μl of the reaction mixture into 5 μl of 500 mm EDTA and heating to 95 °C.

Cell Culture:

Article Title: Role for Bovine Viral Diarrhea Virus Erns Glycoprotein in the Control of Activation of Beta Interferon by Double-Stranded RNA
Article Snippet: The glycoproteins in cell culture medium were further incubated at 37°C for 18 h, and the RNase activity was determined as described previously ( ) with slight modification. .. Briefly, the assay mixture (25 μl in total) contained 0.2 μg of wild-type or mutant Erns , 12.5 μg of yeast 16-23S RNA (Worthington), and 40 U of RNase inhibitor (RNasin; Promega) in 20 mM sodium acetate buffer (pH 4.5).

other:

Article Title: DNA Aptamers to Human Immunodeficiency Virus Reverse Transcriptase Selected by a Primer-Free SELEX Method: Characterization and Comparison with Other Aptamers
Article Snippet: RNase inhibitor (RNasin) was from Promega.

Imaging:

Article Title: Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
Article Snippet: RNase P activity assay RNase P cleavage was performed by mixing 300 nm MRPP1–MRPP2, 150 nm MRPP3, 10 units of RNase inhibitors (RNasin from Promega), and 400 nm in vitro transcribed pre-(mt) tRNAIle in a buffer of 30 mm Tris-HCl, pH 8, 40 mm NaCl, 4.5 mm MgCl2 , and 2 mm DTT to a total reaction volume of 8.25 μl. .. The gel was stained for 30 min with SYBR Gold (Invitrogen) diluted 1000-fold in TBE buffer from the manufacturer's stock and imaged using the Bio-Rad ChemiDoc imaging system.

Sequencing:

Article Title: Codon-usage-based inhibition of HIV protein synthesis by human schlafen 11
Article Snippet: Briefly, freshly harvested cells were gently lysed on ice for 5 min in buffer RLN (50 mM Tris·Cl, pH 8.0, 140 mM NaCl, 1.5 mM MgCl2 , 0.5% (v/v) IGEPAL CA-630, 1,000 U ml−1 RNasin Plus RNase Inhibitor (Promega) and 1 mM DTT). .. RNA was cleaned using Ambion DNA-free kits and reverse transcribed with Applied Biosystems’ High Capacity cDNA Reverse Transcription kit. qPCR was performed on Applied Biosystems 7000 Sequence Detection System or Applied Biosystems StepOnePlus Real-Time PCR System using Power SYBR Green PCR Master Mix.

Article Title: Telomerase in kinetoplastid parasitic protozoa
Article Snippet: The products were fractionated in 10% sequencing gels and autoradiographed or analyzed by PhosphorImager (Molecular Dynamics). .. Activity in the extracts was tested for RNase A sensitivity by incubation with 100 ng of RNase A (Sigma) for 5 min at 37°C before or after the telomerase reaction step and with or without 1 unit of RNase inhibitor RNasin (Promega) before addition of RNase A.

Binding Assay:

Article Title: In Vitro Epsilon RNA-Dependent Protein Priming Activity of Human Hepatitis B Virus Polymerase
Article Snippet: .. After 3 h of binding at room temperature with shaking, unbound materials were removed, and the beads were washed in FLAG lysis buffer with 2 mM DTT, individual protease inhibitors, and 10 U RNasin Plus RNase inhibitor per ml buffer. ..

Molecular Weight:

Article Title: Selective amplification and sequencing of cyclic phosphate-containing RNAs by the cP-RNA-seq method
Article Snippet: Bromophenol Blue (Sigma-Aldrich, cat. no. B0126) Xylene Cyanol FF (Sigma-Aldrich, cat. no. X4126) Low Molecular Weight Marker, 10−100 nt (Affymetrix, cat. no. 76410) SYBR® Gold Nucleic Acid Gel Stain (Life Technologies, cat. no. S-11494) CAUTION! .. Sodium Acetate (3 M), pH 5.5 (Life Technologies, cat. no. AM9740) EDTA (0.5 M), pH 8.0 (Life Technologies, cat. no. AM9260G) SDS, 20% Solution (Life Technologies, cat. no. AM9820) Alkaline Phosphatase, Calf Intestinal (CIP; New England Biolabs Inc, cat. no. M0290) 10× CutSmart Buffer (supplied with CIP) RNasin® Ribonuclease Inhibitor (Promega, cat. no. N2111) Acid-Phenol:Chloroform, pH 4.5 (with IAA, 125:24:1) (Life Technologies, cat. no. AM9720) CAUTION!

Mutagenesis:

Article Title: Role for Bovine Viral Diarrhea Virus Erns Glycoprotein in the Control of Activation of Beta Interferon by Double-Stranded RNA
Article Snippet: .. Briefly, the assay mixture (25 μl in total) contained 0.2 μg of wild-type or mutant Erns , 12.5 μg of yeast 16-23S RNA (Worthington), and 40 U of RNase inhibitor (RNasin; Promega) in 20 mM sodium acetate buffer (pH 4.5). .. The mixture was incubated at 37°C for 45 min, and the enzyme reaction was stopped by acid precipitation with 5 μl of 25% (vol/vol) HClO4 containing 0.75% (wt/vol) uranyl acetate.

Isolation:

Article Title: Listeria monocytogenes ?B Modulates PrfA-Mediated Virulence Factor Expression ▿ Modulates PrfA-Mediated Virulence Factor Expression ▿ †
Article Snippet: RNA isolation was performed using an RNeasy Midi kit (Qiagen, Valencia, CA). .. Samples were treated with DNase for 1 h at 37°C, using 40 units RQ1 DNase enzyme (Promega, Madison, WI) in the presence of 400 units of RNasin Plus RNase inhibitor (Promega).

Article Title: Increased Hematopoietic Extracellular RNAs and Vesicles in the Lung during Allergic Airway Responses
Article Snippet: exRNA stability To test whether synthetic miRNA-like oligoribonucleotide sequences were intrinsically stable in BALF, 22 nucleotide RNA calibrators (calibrators 1-4 ( )) were spiked into BALF and incubated for 5min at 37°C with or without RNAsin ribonuclease inhibitor (Promega) before RNA extraction. .. To test whether endogenous ex-miRNAs were stable to exogenous RNases, RNase A (Thermo Scientific) was added to BALF or total RNA previously isolated from BALF at 0.04-40ug/ml and incubated for 37°C for 15min before RNA extraction.

Article Title: Increased Hematopoietic Extracellular RNAs and Vesicles in the Lung during Allergic Airway Responses
Article Snippet: To test whether synthetic miRNA-like oligoribonucleotide sequences were intrinsically stable in BALF, 22 nucleotide RNA calibrators (calibrators 1-4 ( )) were spiked into BALF and incubated for 5min at 37°C with or without RNAsin ribonuclease inhibitor (Promega) before RNA extraction. .. To test whether endogenous ex-miRNAs were stable to exogenous RNases, RNase A (Thermo Scientific) was added to BALF or total RNA previously isolated from BALF at 0.04-40ug/ml and incubated for 37°C for 15min before RNA extraction.

Microscopy:

Article Title: Zebrafish deficient for Muscleblind-like 2 exhibit features of myotonic dystrophy
Article Snippet: Reactions were set up with 10–40 U of the corresponding (SP6, T7 or T3) RNA polymerase (Roche), 10× concentrated transcription buffer [400 mM Tris-HCl (pH 8.0; 20°C), 60 mM MgCl2 , 100 mM dithiothreitol (DTT), 20 mM spermidine], 20 U RNase inhibitor (RNasin, Promega), 10 mM ATP, 10 mM CTP, 10 mM GTP, 6.5 mM UTP, 3.5 mM DIG-UTP and sterile distilled water to make a total volume of 20 μl. .. The embryos were mounted in glycerol and the results registered using a Nikon digital camera attached to a SMZ1000 Nikon microscope with a cold light source (Photonic).

Purification:

Article Title: Zebrafish deficient for Muscleblind-like 2 exhibit features of myotonic dystrophy
Article Snippet: Whole-mount in situ hybridization Zebrafish mbnl2 plasmid DNA was linearized with Eco RI-Xho I and then purified by phenol/chloroform/isoamyl alcohol extraction and subsequent ethanol precipitation. .. Reactions were set up with 10–40 U of the corresponding (SP6, T7 or T3) RNA polymerase (Roche), 10× concentrated transcription buffer [400 mM Tris-HCl (pH 8.0; 20°C), 60 mM MgCl2 , 100 mM dithiothreitol (DTT), 20 mM spermidine], 20 U RNase inhibitor (RNasin, Promega), 10 mM ATP, 10 mM CTP, 10 mM GTP, 6.5 mM UTP, 3.5 mM DIG-UTP and sterile distilled water to make a total volume of 20 μl.

Article Title: Role for Bovine Viral Diarrhea Virus Erns Glycoprotein in the Control of Activation of Beta Interferon by Double-Stranded RNA
Article Snippet: Purified wild-type and mutant Erns glycoproteins were diluted either in RNase assay buffer (20 mM sodium acetate buffer [pH 4.5]) or cell culture medium (DMEM containing 2% FCS). .. Briefly, the assay mixture (25 μl in total) contained 0.2 μg of wild-type or mutant Erns , 12.5 μg of yeast 16-23S RNA (Worthington), and 40 U of RNase inhibitor (RNasin; Promega) in 20 mM sodium acetate buffer (pH 4.5).

Polymerase Chain Reaction:

Article Title: Codon-usage-based inhibition of HIV protein synthesis by human schlafen 11
Article Snippet: Briefly, freshly harvested cells were gently lysed on ice for 5 min in buffer RLN (50 mM Tris·Cl, pH 8.0, 140 mM NaCl, 1.5 mM MgCl2 , 0.5% (v/v) IGEPAL CA-630, 1,000 U ml−1 RNasin Plus RNase Inhibitor (Promega) and 1 mM DTT). .. RNA was cleaned using Ambion DNA-free kits and reverse transcribed with Applied Biosystems’ High Capacity cDNA Reverse Transcription kit. qPCR was performed on Applied Biosystems 7000 Sequence Detection System or Applied Biosystems StepOnePlus Real-Time PCR System using Power SYBR Green PCR Master Mix.

Article Title: Selective amplification and sequencing of cyclic phosphate-containing RNAs by the cP-RNA-seq method
Article Snippet: Sodium Acetate (3 M), pH 5.5 (Life Technologies, cat. no. AM9740) EDTA (0.5 M), pH 8.0 (Life Technologies, cat. no. AM9260G) SDS, 20% Solution (Life Technologies, cat. no. AM9820) Alkaline Phosphatase, Calf Intestinal (CIP; New England Biolabs Inc, cat. no. M0290) 10× CutSmart Buffer (supplied with CIP) RNasin® Ribonuclease Inhibitor (Promega, cat. no. N2111) Acid-Phenol:Chloroform, pH 4.5 (with IAA, 125:24:1) (Life Technologies, cat. no. AM9720) CAUTION! .. Linear Acrylamide (Life Technologies, cat. no. AM9520) Sodium periodate (Sigma-Aldrich, cat. no. 311448) T4 Polynucleotide Kinase (T4 PNK; New England Biolabs Inc, cat. no. M0201) 10× T4 Polynucleotide Kinase Reaction Buffer (supplied with T4 PNK) Adenosine 5′-Triphosphate (ATP) (New England Biolabs Inc, cat. no. P0756) TruSeq Small RNA Library Preparation Kits (Illumina, cat. no. RS-200-0012), containing RNA 3′ Adapter, Ligation Buffer, Stop Solution, RNA 5′ Adapter, 10 mM ATP, T4 RNA Ligase, 25 mM dNTP mix, RNA RT Primer, PCR Mix, RNA PCR Primer, RNA PCR Primer Index, and High Resolution Ladder.

Article Title: Telomerase in kinetoplastid parasitic protozoa
Article Snippet: The extracts were incubated at 28 °C for 1 h to allow extension of the TS primer by telomerase, followed by PCR amplification using the reverse CX-ext primer, as described ( ). .. Activity in the extracts was tested for RNase A sensitivity by incubation with 100 ng of RNase A (Sigma) for 5 min at 37°C before or after the telomerase reaction step and with or without 1 unit of RNase inhibitor RNasin (Promega) before addition of RNase A.

Labeling:

Article Title: The structure of human Nocturnin reveals a conserved ribonuclease domain that represses target transcript translation and abundance in cells
Article Snippet: .. NOCT64–431 , CNOT6L158–555 or CNOT6L158–555 E240A (1 μM) were mixed with 10 nM 5′ Cy3 labeled p(A)20 (Dharmacon), and RNasin RNase A inhibitor (Promega) and incubated over a 30-min time course at 37°C. ..

Polyacrylamide Gel Electrophoresis:

Article Title: Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
Article Snippet: RNase P activity assay RNase P cleavage was performed by mixing 300 nm MRPP1–MRPP2, 150 nm MRPP3, 10 units of RNase inhibitors (RNasin from Promega), and 400 nm in vitro transcribed pre-(mt) tRNAIle in a buffer of 30 mm Tris-HCl, pH 8, 40 mm NaCl, 4.5 mm MgCl2 , and 2 mm DTT to a total reaction volume of 8.25 μl. .. This sample was analyzed by denaturing PAGE on a 6% TBE-urea gel (Invitrogen) run in TBE buffer (Invitrogen).

Lysis:

Article Title: In Vitro Epsilon RNA-Dependent Protein Priming Activity of Human Hepatitis B Virus Polymerase
Article Snippet: .. After 3 h of binding at room temperature with shaking, unbound materials were removed, and the beads were washed in FLAG lysis buffer with 2 mM DTT, individual protease inhibitors, and 10 U RNasin Plus RNase inhibitor per ml buffer. ..

In Situ Hybridization:

Article Title: Zebrafish deficient for Muscleblind-like 2 exhibit features of myotonic dystrophy
Article Snippet: Paragraph title: Whole-mount in situ hybridization ... Reactions were set up with 10–40 U of the corresponding (SP6, T7 or T3) RNA polymerase (Roche), 10× concentrated transcription buffer [400 mM Tris-HCl (pH 8.0; 20°C), 60 mM MgCl2 , 100 mM dithiothreitol (DTT), 20 mM spermidine], 20 U RNase inhibitor (RNasin, Promega), 10 mM ATP, 10 mM CTP, 10 mM GTP, 6.5 mM UTP, 3.5 mM DIG-UTP and sterile distilled water to make a total volume of 20 μl.

Plasmid Preparation:

Article Title: The RNA Capping Enzyme Domain in Protein A is Essential for Flock House Virus Replication
Article Snippet: Ten µg of T_Rluc plasmid was linearized with Eco RI and electrophoresed in a 1% agarose gel. .. In vitro transcribed uncapped T_Rluc and FHV_T RNA were made by assembling on ice 750 ng linearized DNA, a final concentration of 1 mM (each) ATP/CTP/GTP/UTP, 5 mM dithiothreitol (DTT), transcription buffer (Promega), 0.2 U/µL RNasin ribonuclease inhibitor (Promega) and 40 U T7 RNA polymerase (Promega) in a final volume of 50 µL.

Article Title: Zebrafish deficient for Muscleblind-like 2 exhibit features of myotonic dystrophy
Article Snippet: Sense and antisense riboprobes were obtained by in vitro transcription of 0.2 pmol of linear plasmid DNA. .. Reactions were set up with 10–40 U of the corresponding (SP6, T7 or T3) RNA polymerase (Roche), 10× concentrated transcription buffer [400 mM Tris-HCl (pH 8.0; 20°C), 60 mM MgCl2 , 100 mM dithiothreitol (DTT), 20 mM spermidine], 20 U RNase inhibitor (RNasin, Promega), 10 mM ATP, 10 mM CTP, 10 mM GTP, 6.5 mM UTP, 3.5 mM DIG-UTP and sterile distilled water to make a total volume of 20 μl.

SYBR Green Assay:

Article Title: Codon-usage-based inhibition of HIV protein synthesis by human schlafen 11
Article Snippet: Briefly, freshly harvested cells were gently lysed on ice for 5 min in buffer RLN (50 mM Tris·Cl, pH 8.0, 140 mM NaCl, 1.5 mM MgCl2 , 0.5% (v/v) IGEPAL CA-630, 1,000 U ml−1 RNasin Plus RNase Inhibitor (Promega) and 1 mM DTT). .. RNA was cleaned using Ambion DNA-free kits and reverse transcribed with Applied Biosystems’ High Capacity cDNA Reverse Transcription kit. qPCR was performed on Applied Biosystems 7000 Sequence Detection System or Applied Biosystems StepOnePlus Real-Time PCR System using Power SYBR Green PCR Master Mix.

RNA Extraction:

Article Title: Listeria monocytogenes ?B Modulates PrfA-Mediated Virulence Factor Expression ▿ Modulates PrfA-Mediated Virulence Factor Expression ▿ †
Article Snippet: Paragraph title: RNA extraction. ... Samples were treated with DNase for 1 h at 37°C, using 40 units RQ1 DNase enzyme (Promega, Madison, WI) in the presence of 400 units of RNasin Plus RNase inhibitor (Promega).

Article Title: Increased Hematopoietic Extracellular RNAs and Vesicles in the Lung during Allergic Airway Responses
Article Snippet: .. exRNA stability To test whether synthetic miRNA-like oligoribonucleotide sequences were intrinsically stable in BALF, 22 nucleotide RNA calibrators (calibrators 1-4 ( )) were spiked into BALF and incubated for 5min at 37°C with or without RNAsin ribonuclease inhibitor (Promega) before RNA extraction. .. To test whether endogenous ex-miRNAs were stable to exogenous RNases, RNase A (Thermo Scientific) was added to BALF or total RNA previously isolated from BALF at 0.04-40ug/ml and incubated for 37°C for 15min before RNA extraction.

Article Title: Increased Hematopoietic Extracellular RNAs and Vesicles in the Lung during Allergic Airway Responses
Article Snippet: .. To test whether synthetic miRNA-like oligoribonucleotide sequences were intrinsically stable in BALF, 22 nucleotide RNA calibrators (calibrators 1-4 ( )) were spiked into BALF and incubated for 5min at 37°C with or without RNAsin ribonuclease inhibitor (Promega) before RNA extraction. .. To test whether endogenous ex-miRNAs were stable to exogenous RNases, RNase A (Thermo Scientific) was added to BALF or total RNA previously isolated from BALF at 0.04-40ug/ml and incubated for 37°C for 15min before RNA extraction.

Agarose Gel Electrophoresis:

Article Title: The RNA Capping Enzyme Domain in Protein A is Essential for Flock House Virus Replication
Article Snippet: Ten µg of T_Rluc plasmid was linearized with Eco RI and electrophoresed in a 1% agarose gel. .. In vitro transcribed uncapped T_Rluc and FHV_T RNA were made by assembling on ice 750 ng linearized DNA, a final concentration of 1 mM (each) ATP/CTP/GTP/UTP, 5 mM dithiothreitol (DTT), transcription buffer (Promega), 0.2 U/µL RNasin ribonuclease inhibitor (Promega) and 40 U T7 RNA polymerase (Promega) in a final volume of 50 µL.

Article Title: Listeria monocytogenes ?B Modulates PrfA-Mediated Virulence Factor Expression ▿ Modulates PrfA-Mediated Virulence Factor Expression ▿ †
Article Snippet: Samples were treated with DNase for 1 h at 37°C, using 40 units RQ1 DNase enzyme (Promega, Madison, WI) in the presence of 400 units of RNasin Plus RNase inhibitor (Promega). .. Prior to use, RNA quality was assessed using agarose gel electrophoresis and measurement of 260/280 and 260/230 absorption ratios, using a NanoDrop-1000 instrument (NanoDrop Technologies, Wilmington, DE).

In Vitro:

Article Title: The RNA Capping Enzyme Domain in Protein A is Essential for Flock House Virus Replication
Article Snippet: .. In vitro transcribed uncapped T_Rluc and FHV_T RNA were made by assembling on ice 750 ng linearized DNA, a final concentration of 1 mM (each) ATP/CTP/GTP/UTP, 5 mM dithiothreitol (DTT), transcription buffer (Promega), 0.2 U/µL RNasin ribonuclease inhibitor (Promega) and 40 U T7 RNA polymerase (Promega) in a final volume of 50 µL. .. In vitro transcribed capped T_Rluc and FHV_T RNA were made similarly except for an rNTP mixture comprising 1 mM ATP/CTP/UTP, 0.5 mM GTP and the addition of cap analog m7 G(5′)ppp(5′)G (New England BioLabs, Ipswich, MA, USA) to a final concentration of 1 mM.

Article Title: Zebrafish deficient for Muscleblind-like 2 exhibit features of myotonic dystrophy
Article Snippet: Sense and antisense riboprobes were obtained by in vitro transcription of 0.2 pmol of linear plasmid DNA. .. Reactions were set up with 10–40 U of the corresponding (SP6, T7 or T3) RNA polymerase (Roche), 10× concentrated transcription buffer [400 mM Tris-HCl (pH 8.0; 20°C), 60 mM MgCl2 , 100 mM dithiothreitol (DTT), 20 mM spermidine], 20 U RNase inhibitor (RNasin, Promega), 10 mM ATP, 10 mM CTP, 10 mM GTP, 6.5 mM UTP, 3.5 mM DIG-UTP and sterile distilled water to make a total volume of 20 μl.

Article Title: Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
Article Snippet: .. RNase P activity assay RNase P cleavage was performed by mixing 300 nm MRPP1–MRPP2, 150 nm MRPP3, 10 units of RNase inhibitors (RNasin from Promega), and 400 nm in vitro transcribed pre-(mt) tRNAIle in a buffer of 30 mm Tris-HCl, pH 8, 40 mm NaCl, 4.5 mm MgCl2 , and 2 mm DTT to a total reaction volume of 8.25 μl. .. The reaction was performed at room temperature and stopped at set times by transferring 1 μl of the reaction mixture into 5 μl of 500 mm EDTA and heating to 95 °C.

Ethanol Precipitation:

Article Title: Zebrafish deficient for Muscleblind-like 2 exhibit features of myotonic dystrophy
Article Snippet: Whole-mount in situ hybridization Zebrafish mbnl2 plasmid DNA was linearized with Eco RI-Xho I and then purified by phenol/chloroform/isoamyl alcohol extraction and subsequent ethanol precipitation. .. Reactions were set up with 10–40 U of the corresponding (SP6, T7 or T3) RNA polymerase (Roche), 10× concentrated transcription buffer [400 mM Tris-HCl (pH 8.0; 20°C), 60 mM MgCl2 , 100 mM dithiothreitol (DTT), 20 mM spermidine], 20 U RNase inhibitor (RNasin, Promega), 10 mM ATP, 10 mM CTP, 10 mM GTP, 6.5 mM UTP, 3.5 mM DIG-UTP and sterile distilled water to make a total volume of 20 μl.

Produced:

Article Title: Zebrafish deficient for Muscleblind-like 2 exhibit features of myotonic dystrophy
Article Snippet: A pax2a antisense probe was produced after linearizing the corresponding plasmid cDNA with Bam HI as described elsewhere ( ; ). .. Reactions were set up with 10–40 U of the corresponding (SP6, T7 or T3) RNA polymerase (Roche), 10× concentrated transcription buffer [400 mM Tris-HCl (pH 8.0; 20°C), 60 mM MgCl2 , 100 mM dithiothreitol (DTT), 20 mM spermidine], 20 U RNase inhibitor (RNasin, Promega), 10 mM ATP, 10 mM CTP, 10 mM GTP, 6.5 mM UTP, 3.5 mM DIG-UTP and sterile distilled water to make a total volume of 20 μl.

Concentration Assay:

Article Title: The RNA Capping Enzyme Domain in Protein A is Essential for Flock House Virus Replication
Article Snippet: .. In vitro transcribed uncapped T_Rluc and FHV_T RNA were made by assembling on ice 750 ng linearized DNA, a final concentration of 1 mM (each) ATP/CTP/GTP/UTP, 5 mM dithiothreitol (DTT), transcription buffer (Promega), 0.2 U/µL RNasin ribonuclease inhibitor (Promega) and 40 U T7 RNA polymerase (Promega) in a final volume of 50 µL. .. In vitro transcribed capped T_Rluc and FHV_T RNA were made similarly except for an rNTP mixture comprising 1 mM ATP/CTP/UTP, 0.5 mM GTP and the addition of cap analog m7 G(5′)ppp(5′)G (New England BioLabs, Ipswich, MA, USA) to a final concentration of 1 mM.

Marker:

Article Title: Selective amplification and sequencing of cyclic phosphate-containing RNAs by the cP-RNA-seq method
Article Snippet: Bromophenol Blue (Sigma-Aldrich, cat. no. B0126) Xylene Cyanol FF (Sigma-Aldrich, cat. no. X4126) Low Molecular Weight Marker, 10−100 nt (Affymetrix, cat. no. 76410) SYBR® Gold Nucleic Acid Gel Stain (Life Technologies, cat. no. S-11494) CAUTION! .. Sodium Acetate (3 M), pH 5.5 (Life Technologies, cat. no. AM9740) EDTA (0.5 M), pH 8.0 (Life Technologies, cat. no. AM9260G) SDS, 20% Solution (Life Technologies, cat. no. AM9820) Alkaline Phosphatase, Calf Intestinal (CIP; New England Biolabs Inc, cat. no. M0290) 10× CutSmart Buffer (supplied with CIP) RNasin® Ribonuclease Inhibitor (Promega, cat. no. N2111) Acid-Phenol:Chloroform, pH 4.5 (with IAA, 125:24:1) (Life Technologies, cat. no. AM9720) CAUTION!

Staining:

Article Title: Selective amplification and sequencing of cyclic phosphate-containing RNAs by the cP-RNA-seq method
Article Snippet: Bromophenol Blue (Sigma-Aldrich, cat. no. B0126) Xylene Cyanol FF (Sigma-Aldrich, cat. no. X4126) Low Molecular Weight Marker, 10−100 nt (Affymetrix, cat. no. 76410) SYBR® Gold Nucleic Acid Gel Stain (Life Technologies, cat. no. S-11494) CAUTION! .. Sodium Acetate (3 M), pH 5.5 (Life Technologies, cat. no. AM9740) EDTA (0.5 M), pH 8.0 (Life Technologies, cat. no. AM9260G) SDS, 20% Solution (Life Technologies, cat. no. AM9820) Alkaline Phosphatase, Calf Intestinal (CIP; New England Biolabs Inc, cat. no. M0290) 10× CutSmart Buffer (supplied with CIP) RNasin® Ribonuclease Inhibitor (Promega, cat. no. N2111) Acid-Phenol:Chloroform, pH 4.5 (with IAA, 125:24:1) (Life Technologies, cat. no. AM9720) CAUTION!

Article Title: Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
Article Snippet: RNase P activity assay RNase P cleavage was performed by mixing 300 nm MRPP1–MRPP2, 150 nm MRPP3, 10 units of RNase inhibitors (RNasin from Promega), and 400 nm in vitro transcribed pre-(mt) tRNAIle in a buffer of 30 mm Tris-HCl, pH 8, 40 mm NaCl, 4.5 mm MgCl2 , and 2 mm DTT to a total reaction volume of 8.25 μl. .. The gel was stained for 30 min with SYBR Gold (Invitrogen) diluted 1000-fold in TBE buffer from the manufacturer's stock and imaged using the Bio-Rad ChemiDoc imaging system.

Hood:

Article Title: Selective amplification and sequencing of cyclic phosphate-containing RNAs by the cP-RNA-seq method
Article Snippet: Use a hood, protective clothing, eye protection, and gloves. .. Sodium Acetate (3 M), pH 5.5 (Life Technologies, cat. no. AM9740) EDTA (0.5 M), pH 8.0 (Life Technologies, cat. no. AM9260G) SDS, 20% Solution (Life Technologies, cat. no. AM9820) Alkaline Phosphatase, Calf Intestinal (CIP; New England Biolabs Inc, cat. no. M0290) 10× CutSmart Buffer (supplied with CIP) RNasin® Ribonuclease Inhibitor (Promega, cat. no. N2111) Acid-Phenol:Chloroform, pH 4.5 (with IAA, 125:24:1) (Life Technologies, cat. no. AM9720) CAUTION!

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  • 99
    Promega recombinant rnasin
    2′-F modified <t>siRNA</t> inhibits gene expression in living mice. Representative bioluminescence images of light emitted from living mice transfected, 2 d earlier, with the luciferase expression plasmids pGL3-control. Mice ( N = 4–5 mice per group) received pGL3-control alone ( A ) or were cotransfected with ( B ) 2′-OH GL3 siRNA, ( C ) 2′-OH GL3 siRNA and the RNase inhibitor, <t>RNasin,</t> ( D ) 2′-F GL3 siRNA, ( E ) 2′-F inverted GL2 siRNA. The mouse in panel F received a low pressure i.v. injection of 2′-F GL3 siRNA on day 1 to test if stabilized siRNAs could be taken up by the liver without hydrodynamic transfection. Mice in panels B , C , and D show significant reductions in emitted light as a result of RNA. Hydrodynamic transfection with 2′-F inverted GL2 siRNA and low pressure i.v. transfection of 2′-F GL3 siRNA did not result in significant RNAi.
    Recombinant Rnasin, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 127 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant rnasin/product/Promega
    Average 99 stars, based on 127 article reviews
    Price from $9.99 to $1999.99
    recombinant rnasin - by Bioz Stars, 2020-03
    99/100 stars
      Buy from Supplier

    99
    Promega recombinant rnasin ribonuclease inhibitor
    2′-F modified <t>siRNA</t> inhibits gene expression in living mice. Representative bioluminescence images of light emitted from living mice transfected, 2 d earlier, with the luciferase expression plasmids pGL3-control. Mice ( N = 4–5 mice per group) received pGL3-control alone ( A ) or were cotransfected with ( B ) 2′-OH GL3 siRNA, ( C ) 2′-OH GL3 siRNA and the RNase inhibitor, <t>RNasin,</t> ( D ) 2′-F GL3 siRNA, ( E ) 2′-F inverted GL2 siRNA. The mouse in panel F received a low pressure i.v. injection of 2′-F GL3 siRNA on day 1 to test if stabilized siRNAs could be taken up by the liver without hydrodynamic transfection. Mice in panels B , C , and D show significant reductions in emitted light as a result of RNA. Hydrodynamic transfection with 2′-F inverted GL2 siRNA and low pressure i.v. transfection of 2′-F GL3 siRNA did not result in significant RNAi.
    Recombinant Rnasin Ribonuclease Inhibitor, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 146 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant rnasin ribonuclease inhibitor/product/Promega
    Average 99 stars, based on 146 article reviews
    Price from $9.99 to $1999.99
    recombinant rnasin ribonuclease inhibitor - by Bioz Stars, 2020-03
    99/100 stars
      Buy from Supplier

    Image Search Results


    2′-F modified siRNA inhibits gene expression in living mice. Representative bioluminescence images of light emitted from living mice transfected, 2 d earlier, with the luciferase expression plasmids pGL3-control. Mice ( N = 4–5 mice per group) received pGL3-control alone ( A ) or were cotransfected with ( B ) 2′-OH GL3 siRNA, ( C ) 2′-OH GL3 siRNA and the RNase inhibitor, RNasin, ( D ) 2′-F GL3 siRNA, ( E ) 2′-F inverted GL2 siRNA. The mouse in panel F received a low pressure i.v. injection of 2′-F GL3 siRNA on day 1 to test if stabilized siRNAs could be taken up by the liver without hydrodynamic transfection. Mice in panels B , C , and D show significant reductions in emitted light as a result of RNA. Hydrodynamic transfection with 2′-F inverted GL2 siRNA and low pressure i.v. transfection of 2′-F GL3 siRNA did not result in significant RNAi.

    Journal: RNA

    Article Title: In vivo activity of nuclease-resistant siRNAs

    doi: 10.1261/rna.5239604

    Figure Lengend Snippet: 2′-F modified siRNA inhibits gene expression in living mice. Representative bioluminescence images of light emitted from living mice transfected, 2 d earlier, with the luciferase expression plasmids pGL3-control. Mice ( N = 4–5 mice per group) received pGL3-control alone ( A ) or were cotransfected with ( B ) 2′-OH GL3 siRNA, ( C ) 2′-OH GL3 siRNA and the RNase inhibitor, RNasin, ( D ) 2′-F GL3 siRNA, ( E ) 2′-F inverted GL2 siRNA. The mouse in panel F received a low pressure i.v. injection of 2′-F GL3 siRNA on day 1 to test if stabilized siRNAs could be taken up by the liver without hydrodynamic transfection. Mice in panels B , C , and D show significant reductions in emitted light as a result of RNA. Hydrodynamic transfection with 2′-F inverted GL2 siRNA and low pressure i.v. transfection of 2′-F GL3 siRNA did not result in significant RNAi.

    Article Snippet: Briefly, 1.8 mL of DPBS (with no MgCl2 or CaCl2 ) was mixed with 3 μg of pGL3-Control (Promega), 3 μg of pThAAT , 10 μg of indicated siRNA, and if indicated, 20 μL of recombinant RNasin (Promega).

    Techniques: Modification, Expressing, Mouse Assay, Transfection, Luciferase, Injection