te rnase  (Thermo Fisher)


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    Name:
    TE pH 8 0
    Description:
    Ambion molecular biology grade TE pH 8 0 solution is supplied in one bottle containing 500 mL The buffer is certified RNase free economical and ready to use Due to the ubiquitous presence of RNases manufacturing products for use with RNA is especially challenging Ambion s nuclease free reagents and buffers are manufactured in facilities specifically designed to prevent the introduction of nucleases Highly sensitive RNase assays are performed at several different stages of the manufacturing process to ensure the highest quality These reagents are rigorously tested for contaminating nonspecific endonuclease exonuclease and RNase activity
    Catalog Number:
    am9849
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|DNA Extraction|General gDNA Purification Reagents & Accessories|Genomic DNA Purification
    Category:
    Lab Reagents and Chemicals
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    Structured Review

    Thermo Fisher te rnase
    Ambion molecular biology grade TE pH 8 0 solution is supplied in one bottle containing 500 mL The buffer is certified RNase free economical and ready to use Due to the ubiquitous presence of RNases manufacturing products for use with RNA is especially challenging Ambion s nuclease free reagents and buffers are manufactured in facilities specifically designed to prevent the introduction of nucleases Highly sensitive RNase assays are performed at several different stages of the manufacturing process to ensure the highest quality These reagents are rigorously tested for contaminating nonspecific endonuclease exonuclease and RNase activity
    https://www.bioz.com/result/te rnase/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    te rnase - by Bioz Stars, 2020-07
    90/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Unveiling the Role of the Most Impactful Cardiovascular Risk Locus through Haplotype Editing
    Article Snippet: .. Genomic DNA was extracted by lysis of cell pellets at 55C for ~1hr in tail lysis buffer (100mM Tris HCl (from pH 8.0 stock), 5mM EDTA, 200mM sodium chloride, 0.2% SDS, 290μg/mL proteinase K (Roche Cat. 03115828001)), treatment with RNase A (QIAGEN Cat. 1007885) at room temp. for 15min, separation of aqueous phase by phenol/choloform/isoamyl alcohol (Invitrogen Cat. 15593031) with 20min centrifugation at 4C and 15,000 RCF, precipitation with 3mM sodium acetate (Ambion Cat. AM9740) in 100% ethanol (Fisher Cat. BP2818), cleaning with 70% ethanol, and resuspension in TE buffer (Ambion Cat. AM9849) at 50C for ~1hr. ..

    Selection:

    Article Title: Integrity of chromatin and replicating DNA in nuclei released from fission yeast by semi-automated grinding in liquid nitrogen
    Article Snippet: .. Agarose-gel-electrophoretic size selection of mono-nucleosomal DNA Cleaned, pelleted MNase-digested DNA was dissolved in 300 μl of TE (10 mM Tris-HCl, pH 8.0, 1 mM EDTA, pH 8.0 (nuclease-free, Invitrogen, #AM9858)) per 1 × 1010 -4 × 1010 original cell equivalents, and then quantitated using a NanoDrop-1000 UV spectrophotometer (Nano Drop Technologies, Wilmington DE, USA). .. Up to 55 μg of sample (170 μg/ml), mixed with 1/5 volume of gel-loading solution (0.5% g/ml orange G (sodium salt, Sigma O-7252), 0.25% g/ml xylene cyanol, 15% g/ml Ficoll (type 400, Sigma F-4375), all in TE, pH 8.0) were loaded into an 81-mm × 2-mm × 2.2-3.0-mm well of a 3.5- to 4-mm thick, 1.25% g/ml NuSieve GTG (Lonza #50081) low-melting-temperature-agarose gel in 0.5 × TBE, 0.2 μg/ml ethidium bromide, and electrophoresed between marker ladders (50-bp DNA ladder, Invitrogen; Quanti-ladder, OriGene) at 4 V/cm (electrode separation), 4°C for 4 h. These conditions optimized resolution of nucleosomal DNA bands with minimal diffusional spreading.

    Magnetic Beads:

    Article Title: Balanced chromosomal rearrangements detection by low-pass whole-genome sequencing
    Article Snippet: .. DNA sample 1× TE (pH8.0) (Ambion, Thermo Fisher Scientific, Waltham, MA) 0.2M HCl (Ambion) 0.2M NaOH (Ambion) dNTP Mix (Invitrogen, Thermo Fisher Scientific, Waltham, MA) Biotin-dNTP Mix (Enzymatics, Qiagen Beverly, Beverly, MA) T4 DNA Polymerase (Enzymatics) Klenow DNA Polymerase (Enzymatics) Molecular Biology Agarose (Bio-Rad Laboratories, Hercules, CA) T4 PNK (Enzymatics) 10×T4 DNA Ligase (Enzymatics) 10×T4 Ligase Buffer (Enzymatics) 25mM ATP (Enzymatics) 10×Blue Buffer (Enzymatics) 1 mM dATP (Enzymatics) Klenow3′-5′(exo) (Enzymatics) 2×Rapid Buffer (Enzymatics) PE Adapter Oligo Mix (Invitrogen) PE index Adapter Oligo (Invitrogen) EB buffer (Qiagen, Hilden, Germany) Milli-Q water Anhydrous ethanol Plasmid-safe 10×Buffer (Epicentre Biotechnologies, Illumina, Madison, WI) Plasmid-safeTM DNase (Epicentre Biotechnologies) Exonuclease I (NEB) 0.5 M EDTA (Ambion) DL2000 marker (Tiangen Biotech Co., LTD., Beijing, China) λ-HindIII marker (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, Liaoning, China) 1 kb DNA Ladder (Tiangen) 50 bp DNA Ladder (NEB, Ipswich, MA) Agencourt AMPure beads (Beckman Coulter, Inc., Brea, CA. cat. no. ) Streptavidin Magnetic Beads (Invitrogen) 1.5-ml microcentrifuge tubes (Axygen, Corning, Axygen Scientific Inc, Union City, CA) 50 ml microcentrifuge tubes (BD Biosciences, San Jose, CA) 1.5 ml Non-stick RNase-Free Microfuge Tube (Ambion) Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) .. Hydroshear (Genemachine, San Carlos, CA) Covaris S2 (Covaris, Woburn, MA) NanoDrop 2000™ (Thermo Fisher Scientific) Vortex-5 (Haimen Kylin-Bell Lab Instruments Co., Ltd, Haimen, Jiangsu, China) Magnetic separator (Dexter Magnetic Technologies, Elk Grove Village, IL) Electrophoresis System (Thermo Fisher Scientific) ThermoMixer (Eppendorf, Hamburg, Germany) Microcentrifuge (Eppendorf) Qubit (Thermo Fisher Scientific)

    Spectrophotometry:

    Article Title: Integrity of chromatin and replicating DNA in nuclei released from fission yeast by semi-automated grinding in liquid nitrogen
    Article Snippet: .. Agarose-gel-electrophoretic size selection of mono-nucleosomal DNA Cleaned, pelleted MNase-digested DNA was dissolved in 300 μl of TE (10 mM Tris-HCl, pH 8.0, 1 mM EDTA, pH 8.0 (nuclease-free, Invitrogen, #AM9858)) per 1 × 1010 -4 × 1010 original cell equivalents, and then quantitated using a NanoDrop-1000 UV spectrophotometer (Nano Drop Technologies, Wilmington DE, USA). .. Up to 55 μg of sample (170 μg/ml), mixed with 1/5 volume of gel-loading solution (0.5% g/ml orange G (sodium salt, Sigma O-7252), 0.25% g/ml xylene cyanol, 15% g/ml Ficoll (type 400, Sigma F-4375), all in TE, pH 8.0) were loaded into an 81-mm × 2-mm × 2.2-3.0-mm well of a 3.5- to 4-mm thick, 1.25% g/ml NuSieve GTG (Lonza #50081) low-melting-temperature-agarose gel in 0.5 × TBE, 0.2 μg/ml ethidium bromide, and electrophoresed between marker ladders (50-bp DNA ladder, Invitrogen; Quanti-ladder, OriGene) at 4 V/cm (electrode separation), 4°C for 4 h. These conditions optimized resolution of nucleosomal DNA bands with minimal diffusional spreading.

    Immunoprecipitation:

    Article Title: Long-read ChIA-PET for base-pair resolution mapping of haplotype-specific chromatin interactions
    Article Snippet: .. Monoclonal Antibody against RNA Polymerase II (8WG16), 0.5 ml (Covance, cat. no. MMS-126R) and Polyclonal Anti-CTCF Antibody (Abcam, cat. no. ab70303) Buffer EB (250 ml, Qiagen, cat. no. 19086) Buffer TE (pH8.0, Ambion, cat. no. AM9849) dATP Solution (100 mM) (NEB, cat. no. N0440S) Dimethyl Sulfoxide (DMSO, Sigma, cat. no. D2650-100ML) DNA Clean & Concentrator-5-Capped Columns (200) (Zymo Research, cat. no. D4014) DNA ladder (25 bp, Invitrogen, cat. no. 10597011) DNA LoBind Tubes (1.5 ml, Eppendorf, cat. no. 022431021) dNTPs (10 mM, Life Technologies, cat. no. 18427-088) DPBS, calcium- and magnesium-free (Gibco, cat. no. 14190-250) Dynabeads M-280 Streptavidin (Invitrogen, cat. no.11205D) Dynabeads Protein G for Immunoprecipitation (50 mL) (Life Technologies, cat. no. 10009D) EDTA (pH 8.0, 0.5 M, 500 ml, Ambion, cat. no. AM9261) EGS (Ethylene glycol bis[succinimidylsuccinate], Thermo Fisher Scientific, cat. no. 21565) Formaldehyde (37% [vol/vol], EMD MILLIPORE, cat. no. 344198-250ML) CAUTION: Formaldehyde is toxic; always operate it in a fume hood. .. Glycine (Sigma, cat. no. G8898-500G) GlycoBlue (Life Technologies, cat. no. AM9516) HEPES buffer (1 M, pH7.3, Fisher Scientific, cat. no. BP299-1) I-Block Protein-Based Blocking Reagent (Thermo Fisher, cat. no. T2015) IGEPAL CA-630 (Sigma, cat. no. I8896) Isopropanol (Sigma, cat. no. I-9516-500ml) Klenow Fragment (3'-- > 5' exo-) (NEB, cat. no. M0212L) Maxtract High Density (2 ml, QIAGEN, cat. no. 129056) NaCl Solution (500 ml, 5.0 M, Ambion, cat. no. AM9759) NEB buffer 2 (NEB, cat. no. B7002S) Nextera DNA Sample Prep Kit (Illumina, cat. no. FC-121-1031) Novex precast TBE Gels (4–20% [wt/vol], Invitrogen, cat. no. EC6225BOX) Nuclease-free Water (50 ml, Ambion, cat. no. AM9937) Phenol: Chloroform: IAA (Ambion, cat. no. AM9730) CAUTION Phenol and Chloroform are toxic; operate them in a fume hood.

    Real-time Polymerase Chain Reaction:

    Article Title: A comparative analysis of high-throughput platforms for validation of a circulating microRNA signature in diabetic retinopathy
    Article Snippet: .. Briefly, each sample (100 ng input), underwent 12 PA cycles, then was diluted 1:40 in 0.1X TE (pH 8.0), combined with TaqMan OpenArray PCR Master Mix and loaded onto TaqMan OpenArray Human MicroRNA Panel (AccuFill™ system). qPCR was completed using the QuantStudio 12K Flex (Life Technologies). .. Dynamic Array RT and PA were undertaken with Megaplex RT/PA Primer Pools using the protocol supplied by Fluidigm Corporation.

    Concentration Assay:

    Article Title: Identification of a Chemoreceptor in Pseudomonas aeruginosa That Specifically Mediates Chemotaxis Toward α-Ketoglutarate
    Article Snippet: .. Each 25 μl assay mixture contained 40 μM protein in 5 mM Tris, 5 mM Pipes, 5 mM Mes, pH8.0 and SYPRO orange (Life Technologies) at 5x concentration. ..

    Incubation:

    Article Title: Axonal ribosomes and mRNAs associate with fragile X granules in adult rodent and human brains
    Article Snippet: .. Sections were rinsed three times with PBS and then incubated for 3 h at 37 °C in either TE (10 mM Tris, pH 8.0 + 1 mM EDTA) or TE + RNase (Ambion RNase cocktail @ 1:10; Ambion RNase V1 @ 1:10). .. Tissue was then rinsed three times in PBS, and processed for immunohistochemistry for FXR2P and rRNA as above.

    Polymerase Chain Reaction:

    Article Title: A comparative analysis of high-throughput platforms for validation of a circulating microRNA signature in diabetic retinopathy
    Article Snippet: .. Briefly, each sample (100 ng input), underwent 12 PA cycles, then was diluted 1:40 in 0.1X TE (pH 8.0), combined with TaqMan OpenArray PCR Master Mix and loaded onto TaqMan OpenArray Human MicroRNA Panel (AccuFill™ system). qPCR was completed using the QuantStudio 12K Flex (Life Technologies). .. Dynamic Array RT and PA were undertaken with Megaplex RT/PA Primer Pools using the protocol supplied by Fluidigm Corporation.

    Hood:

    Article Title: Long-read ChIA-PET for base-pair resolution mapping of haplotype-specific chromatin interactions
    Article Snippet: .. Monoclonal Antibody against RNA Polymerase II (8WG16), 0.5 ml (Covance, cat. no. MMS-126R) and Polyclonal Anti-CTCF Antibody (Abcam, cat. no. ab70303) Buffer EB (250 ml, Qiagen, cat. no. 19086) Buffer TE (pH8.0, Ambion, cat. no. AM9849) dATP Solution (100 mM) (NEB, cat. no. N0440S) Dimethyl Sulfoxide (DMSO, Sigma, cat. no. D2650-100ML) DNA Clean & Concentrator-5-Capped Columns (200) (Zymo Research, cat. no. D4014) DNA ladder (25 bp, Invitrogen, cat. no. 10597011) DNA LoBind Tubes (1.5 ml, Eppendorf, cat. no. 022431021) dNTPs (10 mM, Life Technologies, cat. no. 18427-088) DPBS, calcium- and magnesium-free (Gibco, cat. no. 14190-250) Dynabeads M-280 Streptavidin (Invitrogen, cat. no.11205D) Dynabeads Protein G for Immunoprecipitation (50 mL) (Life Technologies, cat. no. 10009D) EDTA (pH 8.0, 0.5 M, 500 ml, Ambion, cat. no. AM9261) EGS (Ethylene glycol bis[succinimidylsuccinate], Thermo Fisher Scientific, cat. no. 21565) Formaldehyde (37% [vol/vol], EMD MILLIPORE, cat. no. 344198-250ML) CAUTION: Formaldehyde is toxic; always operate it in a fume hood. .. Glycine (Sigma, cat. no. G8898-500G) GlycoBlue (Life Technologies, cat. no. AM9516) HEPES buffer (1 M, pH7.3, Fisher Scientific, cat. no. BP299-1) I-Block Protein-Based Blocking Reagent (Thermo Fisher, cat. no. T2015) IGEPAL CA-630 (Sigma, cat. no. I8896) Isopropanol (Sigma, cat. no. I-9516-500ml) Klenow Fragment (3'-- > 5' exo-) (NEB, cat. no. M0212L) Maxtract High Density (2 ml, QIAGEN, cat. no. 129056) NaCl Solution (500 ml, 5.0 M, Ambion, cat. no. AM9759) NEB buffer 2 (NEB, cat. no. B7002S) Nextera DNA Sample Prep Kit (Illumina, cat. no. FC-121-1031) Novex precast TBE Gels (4–20% [wt/vol], Invitrogen, cat. no. EC6225BOX) Nuclease-free Water (50 ml, Ambion, cat. no. AM9937) Phenol: Chloroform: IAA (Ambion, cat. no. AM9730) CAUTION Phenol and Chloroform are toxic; operate them in a fume hood.

    Marker:

    Article Title: Balanced chromosomal rearrangements detection by low-pass whole-genome sequencing
    Article Snippet: .. DNA sample 1× TE (pH8.0) (Ambion, Thermo Fisher Scientific, Waltham, MA) 0.2M HCl (Ambion) 0.2M NaOH (Ambion) dNTP Mix (Invitrogen, Thermo Fisher Scientific, Waltham, MA) Biotin-dNTP Mix (Enzymatics, Qiagen Beverly, Beverly, MA) T4 DNA Polymerase (Enzymatics) Klenow DNA Polymerase (Enzymatics) Molecular Biology Agarose (Bio-Rad Laboratories, Hercules, CA) T4 PNK (Enzymatics) 10×T4 DNA Ligase (Enzymatics) 10×T4 Ligase Buffer (Enzymatics) 25mM ATP (Enzymatics) 10×Blue Buffer (Enzymatics) 1 mM dATP (Enzymatics) Klenow3′-5′(exo) (Enzymatics) 2×Rapid Buffer (Enzymatics) PE Adapter Oligo Mix (Invitrogen) PE index Adapter Oligo (Invitrogen) EB buffer (Qiagen, Hilden, Germany) Milli-Q water Anhydrous ethanol Plasmid-safe 10×Buffer (Epicentre Biotechnologies, Illumina, Madison, WI) Plasmid-safeTM DNase (Epicentre Biotechnologies) Exonuclease I (NEB) 0.5 M EDTA (Ambion) DL2000 marker (Tiangen Biotech Co., LTD., Beijing, China) λ-HindIII marker (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, Liaoning, China) 1 kb DNA Ladder (Tiangen) 50 bp DNA Ladder (NEB, Ipswich, MA) Agencourt AMPure beads (Beckman Coulter, Inc., Brea, CA. cat. no. ) Streptavidin Magnetic Beads (Invitrogen) 1.5-ml microcentrifuge tubes (Axygen, Corning, Axygen Scientific Inc, Union City, CA) 50 ml microcentrifuge tubes (BD Biosciences, San Jose, CA) 1.5 ml Non-stick RNase-Free Microfuge Tube (Ambion) Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) .. Hydroshear (Genemachine, San Carlos, CA) Covaris S2 (Covaris, Woburn, MA) NanoDrop 2000™ (Thermo Fisher Scientific) Vortex-5 (Haimen Kylin-Bell Lab Instruments Co., Ltd, Haimen, Jiangsu, China) Magnetic separator (Dexter Magnetic Technologies, Elk Grove Village, IL) Electrophoresis System (Thermo Fisher Scientific) ThermoMixer (Eppendorf, Hamburg, Germany) Microcentrifuge (Eppendorf) Qubit (Thermo Fisher Scientific)

    Lysis:

    Article Title: Unveiling the Role of the Most Impactful Cardiovascular Risk Locus through Haplotype Editing
    Article Snippet: .. Genomic DNA was extracted by lysis of cell pellets at 55C for ~1hr in tail lysis buffer (100mM Tris HCl (from pH 8.0 stock), 5mM EDTA, 200mM sodium chloride, 0.2% SDS, 290μg/mL proteinase K (Roche Cat. 03115828001)), treatment with RNase A (QIAGEN Cat. 1007885) at room temp. for 15min, separation of aqueous phase by phenol/choloform/isoamyl alcohol (Invitrogen Cat. 15593031) with 20min centrifugation at 4C and 15,000 RCF, precipitation with 3mM sodium acetate (Ambion Cat. AM9740) in 100% ethanol (Fisher Cat. BP2818), cleaning with 70% ethanol, and resuspension in TE buffer (Ambion Cat. AM9849) at 50C for ~1hr. ..

    Methylated DNA Immunoprecipitation:

    Article Title: Comprehensive whole DNA methylome analysis by integrating MeDIP-seq and MRE-seq
    Article Snippet: .. MeDIP elution buffer, mix 12.5 μl (20 mg/ml stock) of proteinase K and 12.5 μl (20% stock) of SDS in a total volume of 1,000 μl (fill up with TE buffer, Life Technologies; cat. no. AM9849). .. Final concentration: 0.25 mg/ml proteinase K and 0.25% SDS in TE.

    Plasmid Preparation:

    Article Title: Balanced chromosomal rearrangements detection by low-pass whole-genome sequencing
    Article Snippet: .. DNA sample 1× TE (pH8.0) (Ambion, Thermo Fisher Scientific, Waltham, MA) 0.2M HCl (Ambion) 0.2M NaOH (Ambion) dNTP Mix (Invitrogen, Thermo Fisher Scientific, Waltham, MA) Biotin-dNTP Mix (Enzymatics, Qiagen Beverly, Beverly, MA) T4 DNA Polymerase (Enzymatics) Klenow DNA Polymerase (Enzymatics) Molecular Biology Agarose (Bio-Rad Laboratories, Hercules, CA) T4 PNK (Enzymatics) 10×T4 DNA Ligase (Enzymatics) 10×T4 Ligase Buffer (Enzymatics) 25mM ATP (Enzymatics) 10×Blue Buffer (Enzymatics) 1 mM dATP (Enzymatics) Klenow3′-5′(exo) (Enzymatics) 2×Rapid Buffer (Enzymatics) PE Adapter Oligo Mix (Invitrogen) PE index Adapter Oligo (Invitrogen) EB buffer (Qiagen, Hilden, Germany) Milli-Q water Anhydrous ethanol Plasmid-safe 10×Buffer (Epicentre Biotechnologies, Illumina, Madison, WI) Plasmid-safeTM DNase (Epicentre Biotechnologies) Exonuclease I (NEB) 0.5 M EDTA (Ambion) DL2000 marker (Tiangen Biotech Co., LTD., Beijing, China) λ-HindIII marker (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, Liaoning, China) 1 kb DNA Ladder (Tiangen) 50 bp DNA Ladder (NEB, Ipswich, MA) Agencourt AMPure beads (Beckman Coulter, Inc., Brea, CA. cat. no. ) Streptavidin Magnetic Beads (Invitrogen) 1.5-ml microcentrifuge tubes (Axygen, Corning, Axygen Scientific Inc, Union City, CA) 50 ml microcentrifuge tubes (BD Biosciences, San Jose, CA) 1.5 ml Non-stick RNase-Free Microfuge Tube (Ambion) Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) .. Hydroshear (Genemachine, San Carlos, CA) Covaris S2 (Covaris, Woburn, MA) NanoDrop 2000™ (Thermo Fisher Scientific) Vortex-5 (Haimen Kylin-Bell Lab Instruments Co., Ltd, Haimen, Jiangsu, China) Magnetic separator (Dexter Magnetic Technologies, Elk Grove Village, IL) Electrophoresis System (Thermo Fisher Scientific) ThermoMixer (Eppendorf, Hamburg, Germany) Microcentrifuge (Eppendorf) Qubit (Thermo Fisher Scientific)

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  • 99
    Thermo Fisher rna
    Individual mRNAs are stabilized under hypoxia in an m 6 A-dependent manner. Total <t>RNA</t> from HEK-293T cells was harvested after 72 h transfection with METTL3/14 siRNA (M3/14 siRNA) or negative control siRNA (Neg siRNA) and 24 h of normoxic or hypoxic conditions and half-life determined via <t>4sU.</t> (#) denotes P ≤ 0.05 by paired Student's t -test between negative siRNA samples in normoxic and hypoxic conditions while (*) denotes P ≤ 0.05 by paired Student's t -test between negative and M3/14 knockdown siRNAs in the hypoxic condition. Error bars represent SEM of five experiments.
    Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3653 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna/product/Thermo Fisher
    Average 99 stars, based on 3653 article reviews
    Price from $9.99 to $1999.99
    rna - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher ercc rna spike in mix
    Quality check of full-length single-cell cDNA libraries. Bioanalyzer profiles of single-cell amplified cDNA from several cells from a typical experiment ( a ) showing variable cDNA yield, as well as a negative control from the same experiment that consisted of <t>ERCC</t> spike-in <t>RNA</t> only (diluted 1:4×10 6 in the RT reaction). Profiles from degraded (b) or contaminated (c) samples are also shown. The low molecular weight fragments in (c) were traced back to bacterial contamination of a particular reagent. All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at Baylor College of Medicine.
    Ercc Rna Spike In Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ercc rna spike in mix/product/Thermo Fisher
    Average 99 stars, based on 39 article reviews
    Price from $9.99 to $1999.99
    ercc rna spike in mix - by Bioz Stars, 2020-07
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      Buy from Supplier

    99
    Thermo Fisher protein a g magnetic beads
    Schematic of the miCLIP protocol. Capped (grey circle) and polyadenylated cellular RNA containing both m 6 Am (red triangle) and m 6 A (red circle) is fragmented and incubated with an anti-m 6 A antibody. Following UV-crosslinking, the antibody-RNA complexes are recovered using protein A/G-affinity beads and a 3’-adapter is ligated to the RNA. Antibody-RNA complexes are then purified by transferring to a nitrocellulose membrane and eluted using proteinase K, leaving only a small peptide fragment crosslinked at the m 6 A/m 6 Am site. RNA fragments are reverse transcribed, which results in mutations or truncations at the crosslink site in the resulting cDNA. Finally, the cDNA is circularized, re-linearized and amplified by PCR to generate final libraries for sequencing.
    Protein A G Magnetic Beads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 361 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protein a g magnetic beads/product/Thermo Fisher
    Average 99 stars, based on 361 article reviews
    Price from $9.99 to $1999.99
    protein a g magnetic beads - by Bioz Stars, 2020-07
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    Image Search Results


    Individual mRNAs are stabilized under hypoxia in an m 6 A-dependent manner. Total RNA from HEK-293T cells was harvested after 72 h transfection with METTL3/14 siRNA (M3/14 siRNA) or negative control siRNA (Neg siRNA) and 24 h of normoxic or hypoxic conditions and half-life determined via 4sU. (#) denotes P ≤ 0.05 by paired Student's t -test between negative siRNA samples in normoxic and hypoxic conditions while (*) denotes P ≤ 0.05 by paired Student's t -test between negative and M3/14 knockdown siRNAs in the hypoxic condition. Error bars represent SEM of five experiments.

    Journal: RNA

    Article Title: N6-methyladenosine is required for the hypoxic stabilization of specific mRNAs

    doi: 10.1261/rna.061044.117

    Figure Lengend Snippet: Individual mRNAs are stabilized under hypoxia in an m 6 A-dependent manner. Total RNA from HEK-293T cells was harvested after 72 h transfection with METTL3/14 siRNA (M3/14 siRNA) or negative control siRNA (Neg siRNA) and 24 h of normoxic or hypoxic conditions and half-life determined via 4sU. (#) denotes P ≤ 0.05 by paired Student's t -test between negative siRNA samples in normoxic and hypoxic conditions while (*) denotes P ≤ 0.05 by paired Student's t -test between negative and M3/14 knockdown siRNAs in the hypoxic condition. Error bars represent SEM of five experiments.

    Article Snippet: Cells were treated with 200 µM 4sU (Sigma-Aldrich) for 1 h. RNA isolated via TRIzol was biotinylated by labeling 50 µg RNA in a reaction mixture with 50 µL 10× Tris/EDTA buffer (TE), 100 µL 1 mg/mL Biotin-HPDP (EZ-Link Biotin HPDP, Thermo Scientific) in dimethylformamide (DMF), and RNase-free H2 O brought to 400 µL.

    Techniques: Transfection, Negative Control

    Quality check of full-length single-cell cDNA libraries. Bioanalyzer profiles of single-cell amplified cDNA from several cells from a typical experiment ( a ) showing variable cDNA yield, as well as a negative control from the same experiment that consisted of ERCC spike-in RNA only (diluted 1:4×10 6 in the RT reaction). Profiles from degraded (b) or contaminated (c) samples are also shown. The low molecular weight fragments in (c) were traced back to bacterial contamination of a particular reagent. All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at Baylor College of Medicine.

    Journal: Nature protocols

    Article Title: Multimodal profiling of single-cell morphology, electrophysiology and gene expression using Patch-seq

    doi: 10.1038/nprot.2017.120

    Figure Lengend Snippet: Quality check of full-length single-cell cDNA libraries. Bioanalyzer profiles of single-cell amplified cDNA from several cells from a typical experiment ( a ) showing variable cDNA yield, as well as a negative control from the same experiment that consisted of ERCC spike-in RNA only (diluted 1:4×10 6 in the RT reaction). Profiles from degraded (b) or contaminated (c) samples are also shown. The low molecular weight fragments in (c) were traced back to bacterial contamination of a particular reagent. All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at Baylor College of Medicine.

    Article Snippet: DNA-OFF (Clontech, cat. no. 9036) Recombinant Ribonuclease Inhibitor (RRI; Clontech, cat. no. 2313A) ERCC RNA spike-in mix (Thermo Fisher Scientific, cat. no. 4456740) Tris-EDTA buffer solution (TE buffer; BioUltra; for molecular biology; Sigma-Aldrich, cat. no. 93283) Triton X-100 (Sigma-Aldrich, cat. no. T8787) CAUTION Harmful if swallowed.

    Techniques: Amplification, Negative Control, Molecular Weight

    Schematic of the miCLIP protocol. Capped (grey circle) and polyadenylated cellular RNA containing both m 6 Am (red triangle) and m 6 A (red circle) is fragmented and incubated with an anti-m 6 A antibody. Following UV-crosslinking, the antibody-RNA complexes are recovered using protein A/G-affinity beads and a 3’-adapter is ligated to the RNA. Antibody-RNA complexes are then purified by transferring to a nitrocellulose membrane and eluted using proteinase K, leaving only a small peptide fragment crosslinked at the m 6 A/m 6 Am site. RNA fragments are reverse transcribed, which results in mutations or truncations at the crosslink site in the resulting cDNA. Finally, the cDNA is circularized, re-linearized and amplified by PCR to generate final libraries for sequencing.

    Journal: Current protocols in molecular biology

    Article Title: Transcriptome-wide mapping of m6A and m6Am at single-nucleotide resolution using miCLIP

    doi: 10.1002/cpmb.88

    Figure Lengend Snippet: Schematic of the miCLIP protocol. Capped (grey circle) and polyadenylated cellular RNA containing both m 6 Am (red triangle) and m 6 A (red circle) is fragmented and incubated with an anti-m 6 A antibody. Following UV-crosslinking, the antibody-RNA complexes are recovered using protein A/G-affinity beads and a 3’-adapter is ligated to the RNA. Antibody-RNA complexes are then purified by transferring to a nitrocellulose membrane and eluted using proteinase K, leaving only a small peptide fragment crosslinked at the m 6 A/m 6 Am site. RNA fragments are reverse transcribed, which results in mutations or truncations at the crosslink site in the resulting cDNA. Finally, the cDNA is circularized, re-linearized and amplified by PCR to generate final libraries for sequencing.

    Article Snippet: TRIzol reagent (Invitrogen 15596018) Oligo d(T)25 magnetic beads (Thermo Scientific 61005 or NEB S1419S) DNase I, RNase-free (Thermo Scientific EN0525) RNA Clean & Concentrator kit (Zymo R1013) 5–20 μg of DNase-treated, poly(A)-selected RNA in 20 μl of water RNA fragmentation reagent, including stop solution (Ambion AM9740) Binding/low-salt buffer (see recipe) Anti-m6 A antibody (such as Abcam ab151230; see Critical Parameters) Protein A/G magnetic beads (Thermo Scientific 88802) High salt buffer (see recipe) PNK wash buffer (see recipe) 5X PNK pH 6.5 buffer (see recipe) T4 polynucleotide kinase (10 U/μl, NEB M0201S) Ribonuclease inhibitor (Promega N2515 or Invitrogen 10777–019) T4 RNA ligase 2, truncated K227Q (200 U/μl, NEB M0351S) L3 linker oligo (20 μM)(see below) PEG-8000 [γ−32 P]ATP, 3000 Ci/mmol (Perkin Elmer BLU002250UC) (see Critical Parameters) 4X LDS sample buffer (Invitrogen NP0008) 1M DTT (Sigma 646563) NuPage Novex 4–12 % bis-tris protein gels (Invitrogen NP0321BOX) MES SDS running buffer (Thermo Scientific NP0002) Prestained protein standard (such as BioRad 161–0373) Bis-Tris transfer buffer (Invitrogen NP0006) Nitrocellulose membrane (0.45 μm pore such as BioRad 1620115) Autoradiography film (such as Carestream Kodak BioMax MS films, Sigma Z363006) SDS Proteinase K (PK) buffer (see recipe) Proteinase K (20 mg/ml, Invitrogen 25530–049) Phase-lock gel heavy tubes (5PRIME 2302830) Acidic phenol:chloroform:ioamyl alcohol (25:24:1, pH 6.5) Nucleic acid co-precipitant (such as GlycoBlue, Ambion AM9516) 2 M sodium chloride 100% ethanol Barcoded reverse transcription (RT) primers (0.5 μM; see below) dNTP mix (10 mM each dNTP) SuperScript III reverse transcriptase (Invitrogen Invitrogen 18080044) 3 M sodium acetate, pH 5.5 2X TBE-urea sample buffer (Invitrogen LC6876) Novex 6% TBE-urea gel (Invitrogen EC6865BOX) Low molecular weight DNA ladder (NEB N3233S) 1X TBE running buffer SYBR Gold Nucleic Acid Stain (Invitrogen S11494) Gel Breaker tubes (IST Engineering 3388–100) 1X Tris-EDTA (TE) buffer CoStar Spin-X columns (0.22 μm pore size, Corning CLS8160) CircLigase II ssDNA ligase (Lucigen CL9025K) FastDigest BamHI (Thermo Scientific FD0054) Cut oligo (10 μM; see below) P3 and P5 PCR primers (10 μM; see below) AccuPrime SuperMix I (Invitrogen 12342010) Novex 6% TBE gel (Invitrogen EC6265BOX) AMPure XP magnetic beads (Beckman Coulter A63880)

    Techniques: Incubation, Purification, Transferring, Amplification, Polymerase Chain Reaction, Sequencing