glycoblue co precipitant  (Thermo Fisher)


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    Name:
    GlycoBlue Coprecipitant 15 mg mL
    Description:
    GlycoBlue Coprecipitant consists of a blue dye covalently linked to glycogen a branched chain carbohydrate which is useful as a nucleic acid coprecipitant The attached dye increases visibility of the pellet This product is an ideal coprecipitant in nuclease protection assays at 1 100 dilution of stock solution Features of GlycoBlue Coprecipitant • Ideal for RT PCR• Increases pellet mass and visibility• Quantitative recovery of low concentrations ng mL of nucleic acid• Prevents pellet loss in nuclease protection assaysWhat is a coprecipitant Coprecipitants are inert substances used to aid recovery of nucleic acids during alcohol precipitations While they can be used for precipitating large amounts of nucleic acids they are essential for quantitative recovery of small amounts of nucleic acids in dilute solutions Often the use of such molecules is desirable for no other reason but visualization of the pelleted precipitate after centrifugation GlycoBlue Coprecipitant can be added to nucleic acid solutions at a final concentration of 50 150 µg mL When a typical acetate alcohol precipitation is done the GlycoBlue Coprecipitant will precipitate with the nucleic acids facilitating good RNA or DNA recovery while increasing the size and visibility of the pellet Since glycogen does not contain appreciable amounts of nucleic acids it is often preferable to yeast RNA as a coprecipitant especially in applications where nucleic acid mass is being assessed or where added nucleic acid could interfere or compete with subsequent enzymatic reactions The glycogen used is isolated from mussel a biological source as are most other preparations of this coprecipitant Glycogen is treated with Proteinase K and SDS to remove any contaminating nucleases and then phenol chloroform extracted ethanol precipitated and resuspended in nuclease free water The glycogen is guaranteed RNase and DNase free
    Catalog Number:
    am9515
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|DNA Extraction|General RNA Purification Reagents & Accessories|General gDNA Purification Reagents & Accessories|Genomic DNA Purification|RNA Extraction|Total RNA Isolation|Total RNA from Animal Cells & Tissues
    Category:
    Lab Reagents and Chemicals
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    Structured Review

    Thermo Fisher glycoblue co precipitant
    GlycoBlue Coprecipitant consists of a blue dye covalently linked to glycogen a branched chain carbohydrate which is useful as a nucleic acid coprecipitant The attached dye increases visibility of the pellet This product is an ideal coprecipitant in nuclease protection assays at 1 100 dilution of stock solution Features of GlycoBlue Coprecipitant • Ideal for RT PCR• Increases pellet mass and visibility• Quantitative recovery of low concentrations ng mL of nucleic acid• Prevents pellet loss in nuclease protection assaysWhat is a coprecipitant Coprecipitants are inert substances used to aid recovery of nucleic acids during alcohol precipitations While they can be used for precipitating large amounts of nucleic acids they are essential for quantitative recovery of small amounts of nucleic acids in dilute solutions Often the use of such molecules is desirable for no other reason but visualization of the pelleted precipitate after centrifugation GlycoBlue Coprecipitant can be added to nucleic acid solutions at a final concentration of 50 150 µg mL When a typical acetate alcohol precipitation is done the GlycoBlue Coprecipitant will precipitate with the nucleic acids facilitating good RNA or DNA recovery while increasing the size and visibility of the pellet Since glycogen does not contain appreciable amounts of nucleic acids it is often preferable to yeast RNA as a coprecipitant especially in applications where nucleic acid mass is being assessed or where added nucleic acid could interfere or compete with subsequent enzymatic reactions The glycogen used is isolated from mussel a biological source as are most other preparations of this coprecipitant Glycogen is treated with Proteinase K and SDS to remove any contaminating nucleases and then phenol chloroform extracted ethanol precipitated and resuspended in nuclease free water The glycogen is guaranteed RNase and DNase free
    https://www.bioz.com/result/glycoblue co precipitant/product/Thermo Fisher
    Average 90 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    glycoblue co precipitant - by Bioz Stars, 2020-02
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    Related Articles

    Centrifugation:

    Article Title: Comprehensive comparative analysis of 5’ end RNA sequencing methods
    Article Snippet: We isolated total RNA using 1 ml Trizol (Thermo Fisher Scientific) according to the manufacturer’s instructions and isopropanol precipitated with 1 μl GlycoBlue Coprecipitant (Thermo Fisher Scientific). .. We removed the GlycoBlue, which inhibited the CAGE process, by mixing with 30 μmol LiCl (Sigma) in a total 12 μl and incubating at −20°C for 20 minutes following by centrifugation at 4°C at maximum speed (17,000 × g).

    Article Title: Assessment of Glyphosate Induced Epigenetic Transgenerational Inheritance of Pathologies and Sperm Epimutations: Generational Toxicology
    Article Snippet: Protein was removed by addition of protein precipitation solution (300 μl, Promega A795A), incubation for 15 min on ice, then centrifugation at 13,500 rpm for 30 minutes at 4 °C. .. One ml of the supernatant was precipitated with 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol.

    Amplification:

    Article Title: Functional signaling and gene regulatory networks between the oocyte and the surrounding cumulus cells
    Article Snippet: We extracted RNA from cumulus cells using the guanidine thiocyanate-phenol chloroform procedure [ , ] (TRIzol reagent, Thermo Fisher), with the addition of 0.5 μl of GlycoBlue Coprecipitant (Thermo Fisher) as the RNA carrier [ ]. .. We eluted the RNA from CCs in 4 μl of a solution containing dNTPs and oligo-dT30 VN and proceeded with polyA+ whole transcriptome amplification using the SMART-seq2 protocol [ , ] for cDNA amplification.

    Article Title: Cholesterol Side-Chain Cleavage Gene Expression in Theca Cells: Augmented Transcriptional Regulation and mRNA Stability in Polycystic Ovary Syndrome
    Article Snippet: Various lengths of the CYP11A1 cDNA were generated by PCR amplification of a pBS-CYP11A1 plasmid containing the full-length (1.86 kbp) CYP11A1 cDNA. .. At each timepoint from 0–60 min, 10 µl of the reaction was removed and immediately precipitated in cold 70% ethanol containing Glycoblue co-precipitant (Ambion, Austin, TX).

    Blocking Assay:

    Article Title: Uncoupling of sgRNAs from their associated barcodes during PCR amplification of combinatorial CRISPR screens
    Article Snippet: When done, turn heat block back on as a token of good lab citizenship, although this will increase the experiment’s carbon footprint. .. Per 50 μL reaction, add in order: 1 μL GlycoBlue (Ambion AM9515) 4 μL NaCl, 5M 55 μL isopropanol Vortex, and incubate at room temperature for 15 minutes.

    Incubation:

    Article Title: A quantitative assay for measuring mRNA decapping by splinted ligation reverse transcription polymerase chain reaction: qSL-RT-PCR
    Article Snippet: Next, 20 U RNase Inhibitor Plus (Promega), 20 U High Concentration T4 DNA ligase (Promega), and T4 DNA ligase buffer were added to each sample and incubated overnight at 15°C. .. The RNA was precipitated with ethanol and 0.3 M sodium acetate (pH 5.2) and 20 μg of glycoblue coprecipitant (Ambion).

    Article Title: Vinclozolin induced epigenetic transgenerational inheritance of pathologies and sperm epimutation biomarkers for specific diseases
    Article Snippet: Then 300 μl of protein precipitation solution (Promega Genomic DNA Purification Kit, A795A) were added, the sample mixed thoroughly and incubated for 15 min on ice. .. One ml of the supernatant was transferred to a 2 ml tube and 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol were added.

    Article Title: The Polycomb-Dependent Epigenome Controls β Cell Dysfunction, Dedifferentiation, and Diabetes
    Article Snippet: .. 500 μL of 2-propanol and 1 μL of GlycoBlue Coprecipitant (Life Technologies) were added to the aqueous phase, mixed and incubated overnight at -20°C before centrifuging them at 12,000 g for 10 min at 4°C. .. Supernatant was decanted and the RNA pellet washed twice with 1 mL 75% ethanol, air-dried and dissolved in 20 μL of RNase free water (Qiagen).

    Article Title: Assessment of Glyphosate Induced Epigenetic Transgenerational Inheritance of Pathologies and Sperm Epimutations: Generational Toxicology
    Article Snippet: Protein was removed by addition of protein precipitation solution (300 μl, Promega A795A), incubation for 15 min on ice, then centrifugation at 13,500 rpm for 30 minutes at 4 °C. .. One ml of the supernatant was precipitated with 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol.

    Modification:

    Article Title: Inter-individual variability in copepod microbiomes reveals bacterial networks linked to host physiology
    Article Snippet: Total genomic DNA was extracted using a modification of a previously described protocol [ ]. .. Subsequently, DNA was precipitated from samples overnight in isopropanol with GlycoBlue (1 µL; Life Technologies #AM9516) as a co-precipitant.

    RNA Binding Assay:

    Article Title: Sensitive PCR-Based Quantitation of Cell-Free Circulating MicroRNAs
    Article Snippet: Centrifuge the samples at 300×g for 15 min at 4° C. Transfer as much as is possible of the clear, upper, aqueous phase without disturbing the interphase, and transfer into new, labeled 1.7 ml microfuge tube (Costar, 3620) or other low RNA-binding tubes are used for extracted RNA). .. Add 1 µl glycoblue (Ambion, AM9515) to each sample.

    Ligation:

    Article Title: Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans
    Article Snippet: The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion). .. Fragmentation efficiency was analyzed with the BioAnalyzer (Agilent), and one-half of each sample (corresponding to ∼100 ng of RNA) was used for adapter ligation. cDNA was gel-purified to capture inserts of RNA fragment size 100–200 nt.

    Methylation:

    Article Title: Assessment of Glyphosate Induced Epigenetic Transgenerational Inheritance of Pathologies and Sperm Epimutations: Generational Toxicology
    Article Snippet: One ml of the supernatant was precipitated with 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol. .. For all generations, equal amounts of DNA from each individual’s sample was used to produce 6 different DNA pools per lineage and the pooled DNA used for methylated DNA immunoprecipitation (MeDIP).

    Cell Culture:

    Article Title: Comprehensive comparative analysis of 5’ end RNA sequencing methods
    Article Snippet: From ScienCell Research Laboratories, we used brain vascular smooth muscle cell RNA (#1105), brain microvascular endothelial cell RNA (#1005), neuron early passage monolayer culture from fetal brain RNA (#1525), and astrocytes cultured for 6 days from a fetal donor RNA (#1805). .. We isolated total RNA using 1 ml Trizol (Thermo Fisher Scientific) according to the manufacturer’s instructions and isopropanol precipitated with 1 μl GlycoBlue Coprecipitant (Thermo Fisher Scientific).

    Generated:

    Article Title: Cholesterol Side-Chain Cleavage Gene Expression in Theca Cells: Augmented Transcriptional Regulation and mRNA Stability in Polycystic Ovary Syndrome
    Article Snippet: Various lengths of the CYP11A1 cDNA were generated by PCR amplification of a pBS-CYP11A1 plasmid containing the full-length (1.86 kbp) CYP11A1 cDNA. .. At each timepoint from 0–60 min, 10 µl of the reaction was removed and immediately precipitated in cold 70% ethanol containing Glycoblue co-precipitant (Ambion, Austin, TX).

    Polymerase Chain Reaction:

    Article Title: Functional signaling and gene regulatory networks between the oocyte and the surrounding cumulus cells
    Article Snippet: We extracted RNA from cumulus cells using the guanidine thiocyanate-phenol chloroform procedure [ , ] (TRIzol reagent, Thermo Fisher), with the addition of 0.5 μl of GlycoBlue Coprecipitant (Thermo Fisher) as the RNA carrier [ ]. .. The samples were subjected to 16 cycles of PCR amplification.

    Article Title: Uncoupling of sgRNAs from their associated barcodes during PCR amplification of combinatorial CRISPR screens
    Article Snippet: Purify by adding to 500 μL buffer PB and proceeding with a PCR spin column purification. .. Per 50 μL reaction, add in order: 1 μL GlycoBlue (Ambion AM9515) 4 μL NaCl, 5M 55 μL isopropanol Vortex, and incubate at room temperature for 15 minutes.

    Article Title: Cholesterol Side-Chain Cleavage Gene Expression in Theca Cells: Augmented Transcriptional Regulation and mRNA Stability in Polycystic Ovary Syndrome
    Article Snippet: The PCR products contained a T7 promoter site and were used to synthesize biotinylated CYP11A1 transcript in the MAXI script kit (Ambion, Austin, TX). .. At each timepoint from 0–60 min, 10 µl of the reaction was removed and immediately precipitated in cold 70% ethanol containing Glycoblue co-precipitant (Ambion, Austin, TX).

    Sonication:

    Article Title: Vinclozolin induced epigenetic transgenerational inheritance of pathologies and sperm epimutation biomarkers for specific diseases
    Article Snippet: The suspension was sonicated for five seconds (Fisher Sonic Dismembrator, model 300, power 25), then centrifuged 5 min at 6000 x g, supernatant discarded and sperm washed again to remove somatic cells and debris. .. One ml of the supernatant was transferred to a 2 ml tube and 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol were added.

    Article Title: Assessment of Glyphosate Induced Epigenetic Transgenerational Inheritance of Pathologies and Sperm Epimutations: Generational Toxicology
    Article Snippet: Somatic cells and debris were therefore removed by brief sonication (Fisher Sonic Dismembrator, model 300, power 25), then centrifugation and washing 1–2 times in 1xPBS. .. One ml of the supernatant was precipitated with 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol.

    Recombinant:

    Article Title: Cholesterol Side-Chain Cleavage Gene Expression in Theca Cells: Augmented Transcriptional Regulation and mRNA Stability in Polycystic Ovary Syndrome
    Article Snippet: Reactions included 0.3 µg of cytoplasmic extract and approximately 10 ng of biotinylated transcript in RNA degradation buffer comprised of 0.6 U recombinant RNAsin (Promega). .. At each timepoint from 0–60 min, 10 µl of the reaction was removed and immediately precipitated in cold 70% ethanol containing Glycoblue co-precipitant (Ambion, Austin, TX).

    Methylated DNA Immunoprecipitation:

    Article Title: Assessment of Glyphosate Induced Epigenetic Transgenerational Inheritance of Pathologies and Sperm Epimutations: Generational Toxicology
    Article Snippet: One ml of the supernatant was precipitated with 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol. .. For all generations, equal amounts of DNA from each individual’s sample was used to produce 6 different DNA pools per lineage and the pooled DNA used for methylated DNA immunoprecipitation (MeDIP).

    DNA Extraction:

    Article Title: Vinclozolin induced epigenetic transgenerational inheritance of pathologies and sperm epimutation biomarkers for specific diseases
    Article Snippet: Paragraph title: Epididymal sperm collection and DNA isolation ... One ml of the supernatant was transferred to a 2 ml tube and 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol were added.

    Article Title: Assessment of Glyphosate Induced Epigenetic Transgenerational Inheritance of Pathologies and Sperm Epimutations: Generational Toxicology
    Article Snippet: Paragraph title: Epididymal sperm collection and DNA isolation ... One ml of the supernatant was precipitated with 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol.

    Article Title: Inter-individual variability in copepod microbiomes reveals bacterial networks linked to host physiology
    Article Snippet: Paragraph title: DNA extraction from individual copepod and seawater samples ... Subsequently, DNA was precipitated from samples overnight in isopropanol with GlycoBlue (1 µL; Life Technologies #AM9516) as a co-precipitant.

    RNA Sequencing Assay:

    Article Title: Dissection of affinity captured LINE-1 macromolecular complexes
    Article Snippet: Paragraph title: RNA sequencing sample preparation and data acquisition ... Aqueous TRIzol extracts were re-extracted in an equal volume of chloroform, and the aqueous phase was again removed; 1 µl (~15 ug) of GlycoBlue (Thermo Fisher Scientific #AM9516) and 2 ul of RNasin were added to this and mixed before combining with 250 µl of isopropanol and incubating for 10’ on ice to precipitate RNA.

    Article Title: Functional signaling and gene regulatory networks between the oocyte and the surrounding cumulus cells
    Article Snippet: Paragraph title: Library preparation and RNA sequencing ... We extracted RNA from cumulus cells using the guanidine thiocyanate-phenol chloroform procedure [ , ] (TRIzol reagent, Thermo Fisher), with the addition of 0.5 μl of GlycoBlue Coprecipitant (Thermo Fisher) as the RNA carrier [ ].

    Article Title: Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans
    Article Snippet: The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion). .. This purified product was then used in the RNase III fragmentation reaction at the beginning of the Solid Total RNA-Seq Kit Whole Transcriptome protocol (Applied Biosystems).

    Magnetic Beads:

    Article Title: Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans
    Article Snippet: Poly-adenylated mRNA was isolated from total RNA using Dynal oligo(dT) magnetic beads (Invitrogen) according to the manufacturer's protocol. .. The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion).

    Isolation:

    Article Title: Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans
    Article Snippet: Poly-adenylated mRNA was isolated from total RNA using Dynal oligo(dT) magnetic beads (Invitrogen) according to the manufacturer's protocol. .. The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion).

    Article Title: Comprehensive comparative analysis of 5’ end RNA sequencing methods
    Article Snippet: .. We isolated total RNA using 1 ml Trizol (Thermo Fisher Scientific) according to the manufacturer’s instructions and isopropanol precipitated with 1 μl GlycoBlue Coprecipitant (Thermo Fisher Scientific). .. We removed the GlycoBlue, which inhibited the CAGE process, by mixing with 30 μmol LiCl (Sigma) in a total 12 μl and incubating at −20°C for 20 minutes following by centrifugation at 4°C at maximum speed (17,000 × g).

    Article Title: The Polycomb-Dependent Epigenome Controls β Cell Dysfunction, Dedifferentiation, and Diabetes
    Article Snippet: Total RNA Extraction Purified islets isolated were lysed directly in TRI reagent and total RNA was extracted according to the manufacturer’s instructions. .. 500 μL of 2-propanol and 1 μL of GlycoBlue Coprecipitant (Life Technologies) were added to the aqueous phase, mixed and incubated overnight at -20°C before centrifuging them at 12,000 g for 10 min at 4°C.

    Clarification Assay:

    Article Title: Dissection of affinity captured LINE-1 macromolecular complexes
    Article Snippet: 600 mg of cell powder was used per preparation, extracted as 3 × 200 mg and pooled after centrifugal clarification, producing ~3 ml of extract. .. Aqueous TRIzol extracts were re-extracted in an equal volume of chloroform, and the aqueous phase was again removed; 1 µl (~15 ug) of GlycoBlue (Thermo Fisher Scientific #AM9516) and 2 ul of RNasin were added to this and mixed before combining with 250 µl of isopropanol and incubating for 10’ on ice to precipitate RNA.

    Labeling:

    Article Title: Sensitive PCR-Based Quantitation of Cell-Free Circulating MicroRNAs
    Article Snippet: Centrifuge the samples at 300×g for 15 min at 4° C. Transfer as much as is possible of the clear, upper, aqueous phase without disturbing the interphase, and transfer into new, labeled 1.7 ml microfuge tube (Costar, 3620) or other low RNA-binding tubes are used for extracted RNA). .. Add 1 µl glycoblue (Ambion, AM9515) to each sample.

    Purification:

    Article Title: Dissection of affinity captured LINE-1 macromolecular complexes
    Article Snippet: Aqueous TRIzol extracts were re-extracted in an equal volume of chloroform, and the aqueous phase was again removed; 1 µl (~15 ug) of GlycoBlue (Thermo Fisher Scientific #AM9516) and 2 ul of RNasin were added to this and mixed before combining with 250 µl of isopropanol and incubating for 10’ on ice to precipitate RNA. .. Extracted RNAs in water were then further purified and concentrated using a Qiagen RNeasy MinElute Cleanup Kit (#74204) following the manufacturer’s instructions, and eluted in 14 µl of nuclease-free water.

    Article Title: Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans
    Article Snippet: .. The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion). .. This purified product was then used in the RNase III fragmentation reaction at the beginning of the Solid Total RNA-Seq Kit Whole Transcriptome protocol (Applied Biosystems).

    Article Title: Uncoupling of sgRNAs from their associated barcodes during PCR amplification of combinatorial CRISPR screens
    Article Snippet: The resulting dsDNA is then ligated into the BsmBI-digested pPapi vector using Golden Gate assembly: 5 μL Tango Buffer (ThermoFisher) 5 μL DTT (stored at -80°C and used once, 10 mM stock) 5 μL ATP (stored at -80°C and used once, 10 mM stock) 500 ng pPapi vector, pre-digested with Esp3I or BsmBI, gel-extracted, and isopropanol-precipitation purified 100 ng dual sgRNA dsDNA insert 1 μL Esp3I (ThermoFisher ER0452) 1 μL T7 ligase (Enzymatics, 3,000 Units / μL L6020L) Up to 50 μL water Cycle 100x (overnight): 5 minutes at 37°C, 5 minutes at 20°C. .. Per 50 μL reaction, add in order: 1 μL GlycoBlue (Ambion AM9515) 4 μL NaCl, 5M 55 μL isopropanol Vortex, and incubate at room temperature for 15 minutes.

    Article Title: The Polycomb-Dependent Epigenome Controls β Cell Dysfunction, Dedifferentiation, and Diabetes
    Article Snippet: Total RNA Extraction Purified islets isolated were lysed directly in TRI reagent and total RNA was extracted according to the manufacturer’s instructions. .. 500 μL of 2-propanol and 1 μL of GlycoBlue Coprecipitant (Life Technologies) were added to the aqueous phase, mixed and incubated overnight at -20°C before centrifuging them at 12,000 g for 10 min at 4°C.

    Lysis:

    Article Title: Functional signaling and gene regulatory networks between the oocyte and the surrounding cumulus cells
    Article Snippet: We extracted RNA from cumulus cells using the guanidine thiocyanate-phenol chloroform procedure [ , ] (TRIzol reagent, Thermo Fisher), with the addition of 0.5 μl of GlycoBlue Coprecipitant (Thermo Fisher) as the RNA carrier [ ]. .. For the oocytes, we added 2 μl of lysis buffer (20 IU/μl RNase inhibitor (Amresco), 0.2% Triton X-100 (Amresco)) to the tube and proceeded with polyA+ whole transcriptome amplification using the SMART-seq2 protocol [ , ] for cDNA amplification.

    Concentration Assay:

    Article Title: A quantitative assay for measuring mRNA decapping by splinted ligation reverse transcription polymerase chain reaction: qSL-RT-PCR
    Article Snippet: Next, 20 U RNase Inhibitor Plus (Promega), 20 U High Concentration T4 DNA ligase (Promega), and T4 DNA ligase buffer were added to each sample and incubated overnight at 15°C. .. The RNA was precipitated with ethanol and 0.3 M sodium acetate (pH 5.2) and 20 μg of glycoblue coprecipitant (Ambion).

    Plasmid Preparation:

    Article Title: Uncoupling of sgRNAs from their associated barcodes during PCR amplification of combinatorial CRISPR screens
    Article Snippet: The resulting dsDNA is then ligated into the BsmBI-digested pPapi vector using Golden Gate assembly: 5 μL Tango Buffer (ThermoFisher) 5 μL DTT (stored at -80°C and used once, 10 mM stock) 5 μL ATP (stored at -80°C and used once, 10 mM stock) 500 ng pPapi vector, pre-digested with Esp3I or BsmBI, gel-extracted, and isopropanol-precipitation purified 100 ng dual sgRNA dsDNA insert 1 μL Esp3I (ThermoFisher ER0452) 1 μL T7 ligase (Enzymatics, 3,000 Units / μL L6020L) Up to 50 μL water Cycle 100x (overnight): 5 minutes at 37°C, 5 minutes at 20°C. .. Per 50 μL reaction, add in order: 1 μL GlycoBlue (Ambion AM9515) 4 μL NaCl, 5M 55 μL isopropanol Vortex, and incubate at room temperature for 15 minutes.

    Article Title: Cholesterol Side-Chain Cleavage Gene Expression in Theca Cells: Augmented Transcriptional Regulation and mRNA Stability in Polycystic Ovary Syndrome
    Article Snippet: Various lengths of the CYP11A1 cDNA were generated by PCR amplification of a pBS-CYP11A1 plasmid containing the full-length (1.86 kbp) CYP11A1 cDNA. .. At each timepoint from 0–60 min, 10 µl of the reaction was removed and immediately precipitated in cold 70% ethanol containing Glycoblue co-precipitant (Ambion, Austin, TX).

    RNA Extraction:

    Article Title: Dissection of affinity captured LINE-1 macromolecular complexes
    Article Snippet: ½ of the sample was saved for RNA extraction (input) and the other ½ was carried forward to split-tandem IP, using 15 µl α-ORF1 affinity medium slurry. .. Aqueous TRIzol extracts were re-extracted in an equal volume of chloroform, and the aqueous phase was again removed; 1 µl (~15 ug) of GlycoBlue (Thermo Fisher Scientific #AM9516) and 2 ul of RNasin were added to this and mixed before combining with 250 µl of isopropanol and incubating for 10’ on ice to precipitate RNA.

    Article Title: Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans
    Article Snippet: Paragraph title: RNA extraction ... The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion).

    Article Title: The Polycomb-Dependent Epigenome Controls β Cell Dysfunction, Dedifferentiation, and Diabetes
    Article Snippet: Paragraph title: Total RNA Extraction ... 500 μL of 2-propanol and 1 μL of GlycoBlue Coprecipitant (Life Technologies) were added to the aqueous phase, mixed and incubated overnight at -20°C before centrifuging them at 12,000 g for 10 min at 4°C.

    Article Title: Neuronal microRNA regulation in Experimental Autoimmune Encephalomyelitis
    Article Snippet: Paragraph title: RNA extraction from LCM tissue ... The upper phase was mixed with 2 times volume of 100% ethanol and 1.5 μl GlycoBlue™ Coprecipitant (ThermoFischer), and left at −80 °C overnight to precipitate all nucleic acids.

    Sample Prep:

    Article Title: Dissection of affinity captured LINE-1 macromolecular complexes
    Article Snippet: Paragraph title: RNA sequencing sample preparation and data acquisition ... Aqueous TRIzol extracts were re-extracted in an equal volume of chloroform, and the aqueous phase was again removed; 1 µl (~15 ug) of GlycoBlue (Thermo Fisher Scientific #AM9516) and 2 ul of RNasin were added to this and mixed before combining with 250 µl of isopropanol and incubating for 10’ on ice to precipitate RNA.

    In Vitro:

    Article Title: Cholesterol Side-Chain Cleavage Gene Expression in Theca Cells: Augmented Transcriptional Regulation and mRNA Stability in Polycystic Ovary Syndrome
    Article Snippet: Paragraph title: In vitro degradation of CYP11A1 mRNA ... At each timepoint from 0–60 min, 10 µl of the reaction was removed and immediately precipitated in cold 70% ethanol containing Glycoblue co-precipitant (Ambion, Austin, TX).

    Homogenization:

    Article Title: Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans
    Article Snippet: One milliliter of TRIzol was used per sample, and homogenization was supplemented with 100 μL of acid-washed sand. .. The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion).

    Ethanol Precipitation:

    Article Title: Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans
    Article Snippet: .. The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion). .. This purified product was then used in the RNase III fragmentation reaction at the beginning of the Solid Total RNA-Seq Kit Whole Transcriptome protocol (Applied Biosystems).

    Laser Capture Microdissection:

    Article Title: Neuronal microRNA regulation in Experimental Autoimmune Encephalomyelitis
    Article Snippet: Paragraph title: RNA extraction from LCM tissue ... The upper phase was mixed with 2 times volume of 100% ethanol and 1.5 μl GlycoBlue™ Coprecipitant (ThermoFischer), and left at −80 °C overnight to precipitate all nucleic acids.

    Immunoprecipitation:

    Article Title: Assessment of Glyphosate Induced Epigenetic Transgenerational Inheritance of Pathologies and Sperm Epimutations: Generational Toxicology
    Article Snippet: One ml of the supernatant was precipitated with 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol. .. For all generations, equal amounts of DNA from each individual’s sample was used to produce 6 different DNA pools per lineage and the pooled DNA used for methylated DNA immunoprecipitation (MeDIP).

    DNA Purification:

    Article Title: Vinclozolin induced epigenetic transgenerational inheritance of pathologies and sperm epimutation biomarkers for specific diseases
    Article Snippet: Then 300 μl of protein precipitation solution (Promega Genomic DNA Purification Kit, A795A) were added, the sample mixed thoroughly and incubated for 15 min on ice. .. One ml of the supernatant was transferred to a 2 ml tube and 2 μl of glycoblue (Invitrogen, AM9516) and 1 ml of cold 100% isopropanol were added.

    Staining:

    Article Title: Genome-Wide Quantitation of Protein Synthesis Rates in Bacteria
    Article Snippet: .. Acid phenol chloroform, pH 4.5 (ThermoFisher, cat. no. AM9720) 20% SDS (ThermoFisher, cat. no. AM9820) Chloroform (Sigma, cat. no. C2432) Sodium Acetate (3 M), pH 5.5, RNase Free (ThermoFisher, cat. no. AM9740) Isopropanol, molecular biology grade (FisherSci, cat. no. BP26181) Ethanol, 200 Proof (VWR, cat. no. AC61509) Tris (1 M), pH 7.0 (FisherSci, cat. no. AM9851) Novex 15% TBE-Urea Gel, 10 well (Thermo, cat. no. EC6885) Novex TBE-Urea Sample Buffer 2× (Thermo, cat. no. LC6876) 10× TBE Buffer (FisherSci, cat. no. AM9863) 10 bp DNA ladder (Invitrogen, cat. no. 10821015) SYBR gold nucleic acid gel stain (Invitrogen, cat. no. S11494) GlycoBlue (Invitrogen, cat. no. AM9516) 5 M NaCl (FisherSci, cat. no. AM9759) T4 Polynucleotide Kinase (NEB, cat. no. M0201L) SUPERase•In™ RNase Inhibitor (TermoFischer, cat. no. AM2696) PEG 8000, 50% (Teknova, cat. No. P4180) T4 RNA ligase 2 truncated K227Q and buffer (NEB, cat. no. M0351S) DEPC-Treated, DNase, RNase free water (Fisher, cat. no. BP561–1) Novex 10% TBE-Urea Gel, 10 well (Thermo, cat. no. EC6875BOX) Deoxynucleotide (dNTP) solution set (NEB, cat. no. N0446S) SuperScript III Reverse Transcriptase (Invitrogen, cat. no. 18080085) DTT (Sigma, cat. no. 10708984001) Sodium hydroxide pellets (Sigma, cat. no. 795429) 1 M Tris pH 8.0 (FisherSci, cat. no. AM9856) 0.5 M EDTA, pH 8.0 (Ambion, cat. no. AM9260G) CircLigase ssDNA Ligase 5000 U (Illumina, cat. no. CL4115K) 20X SSC Buffer, molecular grade (Progmega, cat. no. V4261) Tween-20, reagent grade (VWR, cat. no. 97062–332) Dynabeads® MyOne™ Streptavidin C1 (Thermoscientific, cat. no. 65001) Phusion polymerase (NEB, cat. no. M0530L) 6× DNA loading dye (Thermo, cat. no. R0611) Novex TBE Gels, 8%, 10 well (Invitrogen, cat. no. EC6215BOX) ..

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  • 90
    Thermo Fisher glycoblue co precipitant
    Glycoblue Co Precipitant, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glycoblue co precipitant/product/Thermo Fisher
    Average 90 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    glycoblue co precipitant - by Bioz Stars, 2020-02
    90/100 stars
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