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Promega u rnasin
U Rnasin, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u rnasin - by Bioz Stars, 2020-04
93/100 stars

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Amplification:

Article Title: Activation of Wnt/?-Catenin Signalling Affects Differentiation of Cells Arising from the Cerebellar Ventricular Zone
Article Snippet: The reactions were prepared with 50 ng sample RNA added to 1× RT-Buffer, 10 µM random hexamer primers (Promega), 0.5 mM dNTPs (Invitrogen), 10 u RNAse inhibitor (Promega), 1 µl Sensiscript reverse transcriptase and made up to 20 µl with nuclease free water. .. All primers used had annealing temperatures of ∼58°C and an amplicon size of 90–120 bp.

Quantitative RT-PCR:

Article Title: Genome-wide investigation reveals pathogen-specific and shared signatures in the response of Caenorhabditis elegans to infection
Article Snippet: Paragraph title: qRT-PCR measurements ... This was mixed into a cocktail, 0.5 mM dNTPs (Invitrogen), 1× First Strand Buffer (Invitrogen), 10 mM DTT (Invitrogen), 50 u RNasin (Promega; Madison, Wisconsin, USA) and 400 u SuperScript™ II (Invitrogen).

Article Title: Activation of Wnt/?-Catenin Signalling Affects Differentiation of Cells Arising from the Cerebellar Ventricular Zone
Article Snippet: The reactions were prepared with 50 ng sample RNA added to 1× RT-Buffer, 10 µM random hexamer primers (Promega), 0.5 mM dNTPs (Invitrogen), 10 u RNAse inhibitor (Promega), 1 µl Sensiscript reverse transcriptase and made up to 20 µl with nuclease free water. .. Quantitative RT-PCR (qRT-PCR) was performed using a Precision qPCR SYBRgreen detection kit (PrimerDesign) and commercially synthesised primer pairs (PrimerDesign) with an Opticon DNA Engine (MJ Research) following the manufacturer's instructions.

Real-time Polymerase Chain Reaction:

Article Title: Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Article Snippet: The RT reactions contained 5 µl of sample (1:10 and 1:40 diluted with RT enzyme dilution buffer) or controls, 8 u RNase inhibitor (Promega), 0.32 mM dNTP (Sigma), 360 nM MS2 reverse primer and 8 ng MS2 RNA (Roche) in 25 µl of 1x RT buffer (Promega: 50 mM TRIS-HCl pH 8.3, 5 mM MgCl2 (modified from 3 mM), 75 mM KCl, 10 mM DTT). .. Following the RT reactions (25°C 10 min, 37°C 120 min and 85°C 5 sec), 5 out of 25 µl RT reactions were transferred to optical 96 well plates containing final volume of 25 µl of PCR mix (RNase 2 U [Progema], 1x TaqMan PCR buffer, 1.5 mM MgCl2 , 0.2 mM dNTP, 0.6 µM of each MS2 forward and MS2 reverse primers, 0.2 µM of MS2 probe, 1.25 U of HotStar Taq DNA polymerase [Qiagen]). qPCR reactions were performed at 37°C 30 min, 95°C 15 min, then 95°C 15 sec 60°C 60 sec for 50 cycles using the Chromo4 MJ Research Real time PCR system.

Article Title: Activation of Wnt/?-Catenin Signalling Affects Differentiation of Cells Arising from the Cerebellar Ventricular Zone
Article Snippet: The reactions were prepared with 50 ng sample RNA added to 1× RT-Buffer, 10 µM random hexamer primers (Promega), 0.5 mM dNTPs (Invitrogen), 10 u RNAse inhibitor (Promega), 1 µl Sensiscript reverse transcriptase and made up to 20 µl with nuclease free water. .. Quantitative RT-PCR (qRT-PCR) was performed using a Precision qPCR SYBRgreen detection kit (PrimerDesign) and commercially synthesised primer pairs (PrimerDesign) with an Opticon DNA Engine (MJ Research) following the manufacturer's instructions.

Random Hexamer Labeling:

Article Title: Activation of Wnt/?-Catenin Signalling Affects Differentiation of Cells Arising from the Cerebellar Ventricular Zone
Article Snippet: .. The reactions were prepared with 50 ng sample RNA added to 1× RT-Buffer, 10 µM random hexamer primers (Promega), 0.5 mM dNTPs (Invitrogen), 10 u RNAse inhibitor (Promega), 1 µl Sensiscript reverse transcriptase and made up to 20 µl with nuclease free water. ..

Activity Assay:

Article Title: Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Article Snippet: Paragraph title: TM-PERT assay for reverse transcriptase activity. ... The RT reactions contained 5 µl of sample (1:10 and 1:40 diluted with RT enzyme dilution buffer) or controls, 8 u RNase inhibitor (Promega), 0.32 mM dNTP (Sigma), 360 nM MS2 reverse primer and 8 ng MS2 RNA (Roche) in 25 µl of 1x RT buffer (Promega: 50 mM TRIS-HCl pH 8.3, 5 mM MgCl2 (modified from 3 mM), 75 mM KCl, 10 mM DTT).

Cell Culture:

Article Title: Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Article Snippet: For reverse transcriptase (RT) activity assays, cell culture supernatant fluids were collected from exponentially growing cultures 2–3 d after a medium change. .. The RT reactions contained 5 µl of sample (1:10 and 1:40 diluted with RT enzyme dilution buffer) or controls, 8 u RNase inhibitor (Promega), 0.32 mM dNTP (Sigma), 360 nM MS2 reverse primer and 8 ng MS2 RNA (Roche) in 25 µl of 1x RT buffer (Promega: 50 mM TRIS-HCl pH 8.3, 5 mM MgCl2 (modified from 3 mM), 75 mM KCl, 10 mM DTT).

Article Title: Influence of gag and RRE sequences on HIV-1 RNA packaging signal structure and function
Article Snippet: Virus particles were collected 48 hrs. after transfection by filtering cell culture media through a 0.22 μm filter and centrifuging through a 20% sucrose cushion at 25,000 rpm for 2 hrs. .. Briefly, 2 μl of diluted virus lysate was transferred to Light Cycler 480 96-well plates (Roche) containing PCR mix (10mM Tris, pH 8.3, 50mM KCL, 0.15% Triton X-100, 2mM MgCl2, 0.2mM dNTPs, 1× EvaGreen (Biotium), 1mM DTT, 0.08 μg/ul MS2 RNA, 0.3μM of forward and reverse MS2 primers, 4 u RNasin (Promega) and 0.5 u Go Taq (Promega)).

Article Title: Genome-wide investigation reveals pathogen-specific and shared signatures in the response of Caenorhabditis elegans to infection
Article Snippet: This was mixed into a cocktail, 0.5 mM dNTPs (Invitrogen), 1× First Strand Buffer (Invitrogen), 10 mM DTT (Invitrogen), 50 u RNasin (Promega; Madison, Wisconsin, USA) and 400 u SuperScript™ II (Invitrogen). .. Under 'infected' conditions, C. elegans grown on E. coli OP50 were exposed to pathogenic bacteria at the late-L4 stage, whereas 'control' animals were continuously cultured on E. coli OP50.

Expressing:

Article Title: Genome-wide investigation reveals pathogen-specific and shared signatures in the response of Caenorhabditis elegans to infection
Article Snippet: This was mixed into a cocktail, 0.5 mM dNTPs (Invitrogen), 1× First Strand Buffer (Invitrogen), 10 mM DTT (Invitrogen), 50 u RNasin (Promega; Madison, Wisconsin, USA) and 400 u SuperScript™ II (Invitrogen). .. Expression data were collected as Ct values, where Ct is equal to the number of PCR cycles required to amplify a given gene from a cDNA population.

Article Title: Effect of STK17A on the sensitivity of ovarian cancer cells to paclitaxel and carboplatin
Article Snippet: Cells were transfected with siRNA or pCDNA3flu for 48 h and the level of STK17A expression was measured by a two-step semi-quantitative RT-PCR. .. The reaction was cooled to 4°C, and then 4 µl 5X RT Buffer, 10 mM DTT, 200 u MMLV Reverse Transcriptase and 40 u RNAsin (Promega Corporation) were added resulting in a volume of 20 µl.

Article Title: Activation of Wnt/?-Catenin Signalling Affects Differentiation of Cells Arising from the Cerebellar Ventricular Zone
Article Snippet: Paragraph title: Quantitative gene expression analysis ... The reactions were prepared with 50 ng sample RNA added to 1× RT-Buffer, 10 µM random hexamer primers (Promega), 0.5 mM dNTPs (Invitrogen), 10 u RNAse inhibitor (Promega), 1 µl Sensiscript reverse transcriptase and made up to 20 µl with nuclease free water.

Modification:

Article Title: Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Article Snippet: .. The RT reactions contained 5 µl of sample (1:10 and 1:40 diluted with RT enzyme dilution buffer) or controls, 8 u RNase inhibitor (Promega), 0.32 mM dNTP (Sigma), 360 nM MS2 reverse primer and 8 ng MS2 RNA (Roche) in 25 µl of 1x RT buffer (Promega: 50 mM TRIS-HCl pH 8.3, 5 mM MgCl2 (modified from 3 mM), 75 mM KCl, 10 mM DTT). .. Following the RT reactions (25°C 10 min, 37°C 120 min and 85°C 5 sec), 5 out of 25 µl RT reactions were transferred to optical 96 well plates containing final volume of 25 µl of PCR mix (RNase 2 U [Progema], 1x TaqMan PCR buffer, 1.5 mM MgCl2 , 0.2 mM dNTP, 0.6 µM of each MS2 forward and MS2 reverse primers, 0.2 µM of MS2 probe, 1.25 U of HotStar Taq DNA polymerase [Qiagen]). qPCR reactions were performed at 37°C 30 min, 95°C 15 min, then 95°C 15 sec 60°C 60 sec for 50 cycles using the Chromo4 MJ Research Real time PCR system.

Article Title: Influence of gag and RRE sequences on HIV-1 RNA packaging signal structure and function
Article Snippet: A real-time reverse-transcription PCR assay modified from Vermeire J., et al [ ] was performed to quantify virus content. .. Briefly, 2 μl of diluted virus lysate was transferred to Light Cycler 480 96-well plates (Roche) containing PCR mix (10mM Tris, pH 8.3, 50mM KCL, 0.15% Triton X-100, 2mM MgCl2, 0.2mM dNTPs, 1× EvaGreen (Biotium), 1mM DTT, 0.08 μg/ul MS2 RNA, 0.3μM of forward and reverse MS2 primers, 4 u RNasin (Promega) and 0.5 u Go Taq (Promega)).

Reverse Transcriptase Assay:

Article Title: Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Article Snippet: Reverse transcriptase activities were measured using Taqman fluorogenic 5′-nulcease product-enhanced reverse transcriptase assay (TM-PERT) as described previously in reference and with some modifications ( ). .. The RT reactions contained 5 µl of sample (1:10 and 1:40 diluted with RT enzyme dilution buffer) or controls, 8 u RNase inhibitor (Promega), 0.32 mM dNTP (Sigma), 360 nM MS2 reverse primer and 8 ng MS2 RNA (Roche) in 25 µl of 1x RT buffer (Promega: 50 mM TRIS-HCl pH 8.3, 5 mM MgCl2 (modified from 3 mM), 75 mM KCl, 10 mM DTT).

Transfection:

Article Title: Influence of gag and RRE sequences on HIV-1 RNA packaging signal structure and function
Article Snippet: Virus particles were collected 48 hrs. after transfection by filtering cell culture media through a 0.22 μm filter and centrifuging through a 20% sucrose cushion at 25,000 rpm for 2 hrs. .. Briefly, 2 μl of diluted virus lysate was transferred to Light Cycler 480 96-well plates (Roche) containing PCR mix (10mM Tris, pH 8.3, 50mM KCL, 0.15% Triton X-100, 2mM MgCl2, 0.2mM dNTPs, 1× EvaGreen (Biotium), 1mM DTT, 0.08 μg/ul MS2 RNA, 0.3μM of forward and reverse MS2 primers, 4 u RNasin (Promega) and 0.5 u Go Taq (Promega)).

Article Title: Effect of STK17A on the sensitivity of ovarian cancer cells to paclitaxel and carboplatin
Article Snippet: Cells were transfected with siRNA or pCDNA3flu for 48 h and the level of STK17A expression was measured by a two-step semi-quantitative RT-PCR. .. The reaction was cooled to 4°C, and then 4 µl 5X RT Buffer, 10 mM DTT, 200 u MMLV Reverse Transcriptase and 40 u RNAsin (Promega Corporation) were added resulting in a volume of 20 µl.

Ligation:

Article Title: Nuclear RNA Decay Pathways Aid Rapid Remodeling of Gene Expression in Yeast
Article Snippet: Phosphatase treatment (1x PNK buffer, 8 u TSAP (Promega), 80 u RNasIN (Promega); 37°C for 30 min). .. 3′ linker ligation (1x PNK buffer, 40 u T4 RNA ligase I (NEB), 80 u RNasIN, 1 μM preadenylated 3′ miRCat-33 linker (IDT); 25°C for 6 hr).

Infection:

Article Title: Genome-wide investigation reveals pathogen-specific and shared signatures in the response of Caenorhabditis elegans to infection
Article Snippet: This was mixed into a cocktail, 0.5 mM dNTPs (Invitrogen), 1× First Strand Buffer (Invitrogen), 10 mM DTT (Invitrogen), 50 u RNasin (Promega; Madison, Wisconsin, USA) and 400 u SuperScript™ II (Invitrogen). .. Under 'infected' conditions, C. elegans grown on E. coli OP50 were exposed to pathogenic bacteria at the late-L4 stage, whereas 'control' animals were continuously cultured on E. coli OP50.

Generated:

Article Title: Effect of STK17A on the sensitivity of ovarian cancer cells to paclitaxel and carboplatin
Article Snippet: The reaction was cooled to 4°C, and then 4 µl 5X RT Buffer, 10 mM DTT, 200 u MMLV Reverse Transcriptase and 40 u RNAsin (Promega Corporation) were added resulting in a volume of 20 µl. .. The yield of cDNA was measured according to the STK17A PCR signal generated relative to the internal standard β-actin ( ).

Polymerase Chain Reaction:

Article Title: Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Article Snippet: The RT reactions contained 5 µl of sample (1:10 and 1:40 diluted with RT enzyme dilution buffer) or controls, 8 u RNase inhibitor (Promega), 0.32 mM dNTP (Sigma), 360 nM MS2 reverse primer and 8 ng MS2 RNA (Roche) in 25 µl of 1x RT buffer (Promega: 50 mM TRIS-HCl pH 8.3, 5 mM MgCl2 (modified from 3 mM), 75 mM KCl, 10 mM DTT). .. Following the RT reactions (25°C 10 min, 37°C 120 min and 85°C 5 sec), 5 out of 25 µl RT reactions were transferred to optical 96 well plates containing final volume of 25 µl of PCR mix (RNase 2 U [Progema], 1x TaqMan PCR buffer, 1.5 mM MgCl2 , 0.2 mM dNTP, 0.6 µM of each MS2 forward and MS2 reverse primers, 0.2 µM of MS2 probe, 1.25 U of HotStar Taq DNA polymerase [Qiagen]). qPCR reactions were performed at 37°C 30 min, 95°C 15 min, then 95°C 15 sec 60°C 60 sec for 50 cycles using the Chromo4 MJ Research Real time PCR system.

Article Title: Influence of gag and RRE sequences on HIV-1 RNA packaging signal structure and function
Article Snippet: .. Briefly, 2 μl of diluted virus lysate was transferred to Light Cycler 480 96-well plates (Roche) containing PCR mix (10mM Tris, pH 8.3, 50mM KCL, 0.15% Triton X-100, 2mM MgCl2, 0.2mM dNTPs, 1× EvaGreen (Biotium), 1mM DTT, 0.08 μg/ul MS2 RNA, 0.3μM of forward and reverse MS2 primers, 4 u RNasin (Promega) and 0.5 u Go Taq (Promega)). .. PCR was performed using the Light Cycler 96 (Roche) and the data analyzed with Light Cycler 96 SoftWare 1.1.

Article Title: Genome-wide investigation reveals pathogen-specific and shared signatures in the response of Caenorhabditis elegans to infection
Article Snippet: This was mixed into a cocktail, 0.5 mM dNTPs (Invitrogen), 1× First Strand Buffer (Invitrogen), 10 mM DTT (Invitrogen), 50 u RNasin (Promega; Madison, Wisconsin, USA) and 400 u SuperScript™ II (Invitrogen). .. All qRT-PCRs were carried out using SYBR® PCR Master Mix (Applied Biosystems; Foster City, California, USA) according to manufacturer's specifications and analyzed on a GeneAmp® 5700 (Perkin Elmer).

Article Title: The CCA anticodon specifies separate functions inside and outside translation in Bacillus cereus
Article Snippet: .. The mixture was heated at 80°C for 2 min followed by incubation at 50°C for 15 min. To the mixture were added 10 mM DTT, first strand buffer, 200 u Superscript Reverse Transcriptase II (Invitrogen) and 20 u RNAsin (Promega), followed by incubation at 50°C for 35 min. PCR was performed upon addition of 3′-end oligonucleotide (1 μM), 5′ end oligonucleotide (2 μM), Pfu DNA polymerase buffer (20 mM Tris HCl, pH 8; 10 mM (NH4 )2 SO4 ; 2 mM MgSO4 ; 0.1% triton X100; 0.1 mg/ml BSA, 10 mM KCl), 0.3 mM dNTPs and 5 u Pfu DNA polymerase (Stratagene) to the RT mixture, followed by 25 cycles of 94°C for 30 s, 55°C for 1 min and 72°C for 1 min, and 1 cycle at 72°C for 10 min. .. The product of RT-PCR was resolved on a 2.5% agarose gel and visualized by staining with ethidium bromide.

Article Title: Effect of STK17A on the sensitivity of ovarian cancer cells to paclitaxel and carboplatin
Article Snippet: The reaction was cooled to 4°C, and then 4 µl 5X RT Buffer, 10 mM DTT, 200 u MMLV Reverse Transcriptase and 40 u RNAsin (Promega Corporation) were added resulting in a volume of 20 µl. .. The yield of cDNA was measured according to the STK17A PCR signal generated relative to the internal standard β-actin ( ).

Article Title: Activation of Wnt/?-Catenin Signalling Affects Differentiation of Cells Arising from the Cerebellar Ventricular Zone
Article Snippet: Quantitative gene expression analysis RNA extraction for reverse transcriptase PCR (RT-PCR) was performed on individual snap frozen cerebellum slices using an RNeasy Microkit (Qiagen) following the manufacturer's instructions. .. The reactions were prepared with 50 ng sample RNA added to 1× RT-Buffer, 10 µM random hexamer primers (Promega), 0.5 mM dNTPs (Invitrogen), 10 u RNAse inhibitor (Promega), 1 µl Sensiscript reverse transcriptase and made up to 20 µl with nuclease free water.

Affinity Purification:

Article Title: Nuclear RNA Decay Pathways Aid Rapid Remodeling of Gene Expression in Yeast
Article Snippet: The solution was adjusted for nickel affinity purification with the addition of 27 μL NaCl (5.0 M) and 3 μL imidazole (2.5 M), and added to 50 μl washed nickel beads (Ni-NTA agarose, QIAGEN). .. Phosphatase treatment (1x PNK buffer, 8 u TSAP (Promega), 80 u RNasIN (Promega); 37°C for 30 min).

Radioactivity:

Article Title: Nuclear RNA Decay Pathways Aid Rapid Remodeling of Gene Expression in Yeast
Article Snippet: Phosphatase treatment (1x PNK buffer, 8 u TSAP (Promega), 80 u RNasIN (Promega); 37°C for 30 min). .. 5′ end phosphorylation and radiolabeling (1x PNK buffer, 40 u T4 PNK (NEB), 40μCi 32 P-γATP; 37°C for 60 min, with addition of 100 nmol of ATP after 40 min).

Size-exclusion Chromatography:

Article Title: Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Article Snippet: The RT reactions contained 5 µl of sample (1:10 and 1:40 diluted with RT enzyme dilution buffer) or controls, 8 u RNase inhibitor (Promega), 0.32 mM dNTP (Sigma), 360 nM MS2 reverse primer and 8 ng MS2 RNA (Roche) in 25 µl of 1x RT buffer (Promega: 50 mM TRIS-HCl pH 8.3, 5 mM MgCl2 (modified from 3 mM), 75 mM KCl, 10 mM DTT). .. Following the RT reactions (25°C 10 min, 37°C 120 min and 85°C 5 sec), 5 out of 25 µl RT reactions were transferred to optical 96 well plates containing final volume of 25 µl of PCR mix (RNase 2 U [Progema], 1x TaqMan PCR buffer, 1.5 mM MgCl2 , 0.2 mM dNTP, 0.6 µM of each MS2 forward and MS2 reverse primers, 0.2 µM of MS2 probe, 1.25 U of HotStar Taq DNA polymerase [Qiagen]). qPCR reactions were performed at 37°C 30 min, 95°C 15 min, then 95°C 15 sec 60°C 60 sec for 50 cycles using the Chromo4 MJ Research Real time PCR system.

Reverse Transcription Polymerase Chain Reaction:

Article Title: The CCA anticodon specifies separate functions inside and outside translation in Bacillus cereus
Article Snippet: Paragraph title: RNA extraction and RT-PCR ... The mixture was heated at 80°C for 2 min followed by incubation at 50°C for 15 min. To the mixture were added 10 mM DTT, first strand buffer, 200 u Superscript Reverse Transcriptase II (Invitrogen) and 20 u RNAsin (Promega), followed by incubation at 50°C for 35 min. PCR was performed upon addition of 3′-end oligonucleotide (1 μM), 5′ end oligonucleotide (2 μM), Pfu DNA polymerase buffer (20 mM Tris HCl, pH 8; 10 mM (NH4 )2 SO4 ; 2 mM MgSO4 ; 0.1% triton X100; 0.1 mg/ml BSA, 10 mM KCl), 0.3 mM dNTPs and 5 u Pfu DNA polymerase (Stratagene) to the RT mixture, followed by 25 cycles of 94°C for 30 s, 55°C for 1 min and 72°C for 1 min, and 1 cycle at 72°C for 10 min.

Article Title: Effect of STK17A on the sensitivity of ovarian cancer cells to paclitaxel and carboplatin
Article Snippet: Paragraph title: Semi-quantitative RT-PCR analysis ... The reaction was cooled to 4°C, and then 4 µl 5X RT Buffer, 10 mM DTT, 200 u MMLV Reverse Transcriptase and 40 u RNAsin (Promega Corporation) were added resulting in a volume of 20 µl.

Article Title: Activation of Wnt/?-Catenin Signalling Affects Differentiation of Cells Arising from the Cerebellar Ventricular Zone
Article Snippet: Quantitative gene expression analysis RNA extraction for reverse transcriptase PCR (RT-PCR) was performed on individual snap frozen cerebellum slices using an RNeasy Microkit (Qiagen) following the manufacturer's instructions. .. The reactions were prepared with 50 ng sample RNA added to 1× RT-Buffer, 10 µM random hexamer primers (Promega), 0.5 mM dNTPs (Invitrogen), 10 u RNAse inhibitor (Promega), 1 µl Sensiscript reverse transcriptase and made up to 20 µl with nuclease free water.

Software:

Article Title: Influence of gag and RRE sequences on HIV-1 RNA packaging signal structure and function
Article Snippet: Briefly, 2 μl of diluted virus lysate was transferred to Light Cycler 480 96-well plates (Roche) containing PCR mix (10mM Tris, pH 8.3, 50mM KCL, 0.15% Triton X-100, 2mM MgCl2, 0.2mM dNTPs, 1× EvaGreen (Biotium), 1mM DTT, 0.08 μg/ul MS2 RNA, 0.3μM of forward and reverse MS2 primers, 4 u RNasin (Promega) and 0.5 u Go Taq (Promega)). .. PCR was performed using the Light Cycler 96 (Roche) and the data analyzed with Light Cycler 96 SoftWare 1.1.

RNA Extraction:

Article Title: The CCA anticodon specifies separate functions inside and outside translation in Bacillus cereus
Article Snippet: Paragraph title: RNA extraction and RT-PCR ... The mixture was heated at 80°C for 2 min followed by incubation at 50°C for 15 min. To the mixture were added 10 mM DTT, first strand buffer, 200 u Superscript Reverse Transcriptase II (Invitrogen) and 20 u RNAsin (Promega), followed by incubation at 50°C for 35 min. PCR was performed upon addition of 3′-end oligonucleotide (1 μM), 5′ end oligonucleotide (2 μM), Pfu DNA polymerase buffer (20 mM Tris HCl, pH 8; 10 mM (NH4 )2 SO4 ; 2 mM MgSO4 ; 0.1% triton X100; 0.1 mg/ml BSA, 10 mM KCl), 0.3 mM dNTPs and 5 u Pfu DNA polymerase (Stratagene) to the RT mixture, followed by 25 cycles of 94°C for 30 s, 55°C for 1 min and 72°C for 1 min, and 1 cycle at 72°C for 10 min.

Article Title: Activation of Wnt/?-Catenin Signalling Affects Differentiation of Cells Arising from the Cerebellar Ventricular Zone
Article Snippet: Quantitative gene expression analysis RNA extraction for reverse transcriptase PCR (RT-PCR) was performed on individual snap frozen cerebellum slices using an RNeasy Microkit (Qiagen) following the manufacturer's instructions. .. The reactions were prepared with 50 ng sample RNA added to 1× RT-Buffer, 10 µM random hexamer primers (Promega), 0.5 mM dNTPs (Invitrogen), 10 u RNAse inhibitor (Promega), 1 µl Sensiscript reverse transcriptase and made up to 20 µl with nuclease free water.

Agarose Gel Electrophoresis:

Article Title: The CCA anticodon specifies separate functions inside and outside translation in Bacillus cereus
Article Snippet: The mixture was heated at 80°C for 2 min followed by incubation at 50°C for 15 min. To the mixture were added 10 mM DTT, first strand buffer, 200 u Superscript Reverse Transcriptase II (Invitrogen) and 20 u RNAsin (Promega), followed by incubation at 50°C for 35 min. PCR was performed upon addition of 3′-end oligonucleotide (1 μM), 5′ end oligonucleotide (2 μM), Pfu DNA polymerase buffer (20 mM Tris HCl, pH 8; 10 mM (NH4 )2 SO4 ; 2 mM MgSO4 ; 0.1% triton X100; 0.1 mg/ml BSA, 10 mM KCl), 0.3 mM dNTPs and 5 u Pfu DNA polymerase (Stratagene) to the RT mixture, followed by 25 cycles of 94°C for 30 s, 55°C for 1 min and 72°C for 1 min, and 1 cycle at 72°C for 10 min. .. The product of RT-PCR was resolved on a 2.5% agarose gel and visualized by staining with ethidium bromide.

Incubation:

Article Title: Genome-wide investigation reveals pathogen-specific and shared signatures in the response of Caenorhabditis elegans to infection
Article Snippet: Total RNA (2.5 μg) was mixed with 2.5 μg (dT)24 -V primer, incubated at 70°C for 10 minutes, then cooled on ice for 5 minutes. .. This was mixed into a cocktail, 0.5 mM dNTPs (Invitrogen), 1× First Strand Buffer (Invitrogen), 10 mM DTT (Invitrogen), 50 u RNasin (Promega; Madison, Wisconsin, USA) and 400 u SuperScript™ II (Invitrogen).

Article Title: The CCA anticodon specifies separate functions inside and outside translation in Bacillus cereus
Article Snippet: .. The mixture was heated at 80°C for 2 min followed by incubation at 50°C for 15 min. To the mixture were added 10 mM DTT, first strand buffer, 200 u Superscript Reverse Transcriptase II (Invitrogen) and 20 u RNAsin (Promega), followed by incubation at 50°C for 35 min. PCR was performed upon addition of 3′-end oligonucleotide (1 μM), 5′ end oligonucleotide (2 μM), Pfu DNA polymerase buffer (20 mM Tris HCl, pH 8; 10 mM (NH4 )2 SO4 ; 2 mM MgSO4 ; 0.1% triton X100; 0.1 mg/ml BSA, 10 mM KCl), 0.3 mM dNTPs and 5 u Pfu DNA polymerase (Stratagene) to the RT mixture, followed by 25 cycles of 94°C for 30 s, 55°C for 1 min and 72°C for 1 min, and 1 cycle at 72°C for 10 min. .. The product of RT-PCR was resolved on a 2.5% agarose gel and visualized by staining with ethidium bromide.

Article Title: Nuclear RNA Decay Pathways Aid Rapid Remodeling of Gene Expression in Yeast
Article Snippet: Following an overnight incubation, the nickel beads were transferred to a spin column and washed three times with WBI (6.0 M guanidine hydrochloride, 50 mM Tris-HCl pH = 7.8, 300 mM NaCl, 0.1% NP-40, 10 mM imidazole, and 5 mM β-mercaptoethanol), and then three times with 1xPNK buffer (50mM Tris-HCl pH = 7.8, 10 mM MgCl2 , 0.5% NP-40, and 5 mM β-mercaptoethanol). .. Phosphatase treatment (1x PNK buffer, 8 u TSAP (Promega), 80 u RNasIN (Promega); 37°C for 30 min).

Virus Isolation Assay:

Article Title: Influence of gag and RRE sequences on HIV-1 RNA packaging signal structure and function
Article Snippet: Paragraph title: Virus isolation & quantification ... Briefly, 2 μl of diluted virus lysate was transferred to Light Cycler 480 96-well plates (Roche) containing PCR mix (10mM Tris, pH 8.3, 50mM KCL, 0.15% Triton X-100, 2mM MgCl2, 0.2mM dNTPs, 1× EvaGreen (Biotium), 1mM DTT, 0.08 μg/ul MS2 RNA, 0.3μM of forward and reverse MS2 primers, 4 u RNasin (Promega) and 0.5 u Go Taq (Promega)).

Standard Deviation:

Article Title: Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Article Snippet: The RT reactions contained 5 µl of sample (1:10 and 1:40 diluted with RT enzyme dilution buffer) or controls, 8 u RNase inhibitor (Promega), 0.32 mM dNTP (Sigma), 360 nM MS2 reverse primer and 8 ng MS2 RNA (Roche) in 25 µl of 1x RT buffer (Promega: 50 mM TRIS-HCl pH 8.3, 5 mM MgCl2 (modified from 3 mM), 75 mM KCl, 10 mM DTT). .. Under our defined conditions , the positive threshold value was set as ≥ 94.6 nU/µL, which was determined as a sum of mean background value (29.9 nU/µL) of culture supernatant fluids from nine different MLV negative cell lines plus three times of Standard Deviation (21.6 nU/µL) as described previously in reference and .

Staining:

Article Title: The CCA anticodon specifies separate functions inside and outside translation in Bacillus cereus
Article Snippet: The mixture was heated at 80°C for 2 min followed by incubation at 50°C for 15 min. To the mixture were added 10 mM DTT, first strand buffer, 200 u Superscript Reverse Transcriptase II (Invitrogen) and 20 u RNAsin (Promega), followed by incubation at 50°C for 35 min. PCR was performed upon addition of 3′-end oligonucleotide (1 μM), 5′ end oligonucleotide (2 μM), Pfu DNA polymerase buffer (20 mM Tris HCl, pH 8; 10 mM (NH4 )2 SO4 ; 2 mM MgSO4 ; 0.1% triton X100; 0.1 mg/ml BSA, 10 mM KCl), 0.3 mM dNTPs and 5 u Pfu DNA polymerase (Stratagene) to the RT mixture, followed by 25 cycles of 94°C for 30 s, 55°C for 1 min and 72°C for 1 min, and 1 cycle at 72°C for 10 min. .. The product of RT-PCR was resolved on a 2.5% agarose gel and visualized by staining with ethidium bromide.

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    u recombinant rnasin ribonuclease inhibitor - by Bioz Stars, 2020-04
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    Promega u reaction rnasin
    U Reaction Rnasin, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/u reaction rnasin/product/Promega
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    u reaction rnasin - by Bioz Stars, 2020-04
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