trizol ls reagent  (Thermo Fisher)


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    Thermo Fisher trizol ls reagent
    Trizol Ls Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 435 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trizol ls reagent/product/Thermo Fisher
    Average 99 stars, based on 435 article reviews
    Price from $9.99 to $1999.99
    trizol ls reagent - by Bioz Stars, 2020-05
    99/100 stars

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    Real-time Polymerase Chain Reaction:

    Article Title: Cell-specific exon methylation and CTCF binding in neurons regulate calcium ion channel splicing and function
    Article Snippet: .. RNA extraction, RT-PCR and qPCR RNA extraction from F11 cells was performed using TRIzol (Invitrogen Cat# 15596018) and from dissociated DRG cells using TRIzol LS Reagent (Invitrogen Cat# 10296010) according to the manufacturer’s instructions. .. 1–2 μg of RNA was immediately reverse transcribed to cDNA using the SuperScript III First-Strand Synthesis System with Poli-dT primers (Invitrogen Cat# 18080051).

    RNA Extraction:

    Article Title: Investigation of a large waterborne acute gastroenteritis outbreak caused by group B rotavirus in Maharashtra state, India, et al. Investigation of a large waterborne acute gastroenteritis outbreak caused by Group B rotavirus in Maharashtra state, India
    Article Snippet: .. 2.3 RNA extraction and viral RNA electrophoresis Viral RNA was extracted from the supernatant of 30% fecal suspensions using TRIzol LS reagent (Invitrogen, Waltham, MA) according to the manufacturer's instructions. .. RNA polyacrylamide gel electrophoresis (RNA‐PAGE) was used for detection of rotavirus double‐stranded RNA genome segments.

    Article Title: Cell-specific exon methylation and CTCF binding in neurons regulate calcium ion channel splicing and function
    Article Snippet: .. RNA extraction, RT-PCR and qPCR RNA extraction from F11 cells was performed using TRIzol (Invitrogen Cat# 15596018) and from dissociated DRG cells using TRIzol LS Reagent (Invitrogen Cat# 10296010) according to the manufacturer’s instructions. .. 1–2 μg of RNA was immediately reverse transcribed to cDNA using the SuperScript III First-Strand Synthesis System with Poli-dT primers (Invitrogen Cat# 18080051).

    Article Title: An Infectious cDNA Clone of SARS-CoV-2
    Article Snippet: .. RNA Extraction, RT-PCR and Sanger Sequencing 250 μL of culture fluids were mixed with three volumes of TRIzol LS Reagent (Thermo Fisher Scientific). ..

    Isolation:

    Article Title: Direct comparison of size-dependent versus EpCAM-dependent CTC enrichment at the gene expression and DNA methylation level in head and neck squamous cell carcinoma
    Article Snippet: .. gDNA isolation from CTCs gDNA was extracted from CTCs using the TRIZOL-LS reagent (ThermoFisher Scientific, USA) as previously described. .. Isolated gDNA was dissolved in 30 μL of 8 mmol/L NaOH.

    Article Title: Translational derepression of Elavl4 isoforms at their alternative 5′ UTRs determines neuronal development
    Article Snippet: .. Briefly, RNA was isolated from input and polysome fractions by using TRIzol LS (Life Technologies; no. 10296028) following the manufacturer’s protocol. ..

    Article Title: Innate Immune Responses of Skin Mucosa in Common Carp (Cyprinus Carpio) Fed a Diet Supplemented with Galactooligosaccharides
    Article Snippet: .. Prior to total RNA isolation, the samples of skin mucosa were homogenized with the TissueRuptor homogenizer (Qiagen GmbH, Hilden, Germany) in TRIzol® LS Reagent (Ambion/Thermo Fisher Scientific, Waltham, MA, USA). .. The lysate was processed using a EURx Universal RNA Purification Kit (EURx, Gdansk, Poland).

    Electrophoresis:

    Article Title: Investigation of a large waterborne acute gastroenteritis outbreak caused by group B rotavirus in Maharashtra state, India, et al. Investigation of a large waterborne acute gastroenteritis outbreak caused by Group B rotavirus in Maharashtra state, India
    Article Snippet: .. 2.3 RNA extraction and viral RNA electrophoresis Viral RNA was extracted from the supernatant of 30% fecal suspensions using TRIzol LS reagent (Invitrogen, Waltham, MA) according to the manufacturer's instructions. .. RNA polyacrylamide gel electrophoresis (RNA‐PAGE) was used for detection of rotavirus double‐stranded RNA genome segments.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: In vitro inhibition of CSFV replication by retroviral vector-mediated RNA interference
    Article Snippet: .. 2.5.2 Construction of 5′-UTR plasmid Total RNA of CSFV-infected cells was extracted with Trizol LS Reagent (Invitrogen, Carlsbad, CA, USA) and then applied as a template for synthesis of the first cDNA strand of the 5′-UTR by RT-PCR using AMV reverse transcriptase (Promega, Madison, WI, USA) and 5′-UTR RP267 primer, according to the manufacturer's instructions. .. PCR was then used to amplify the 5′-UTR with FP147 and RP267 primers under conditions of 94 °C for 2 min, then 35 cycles of 94 °C for 30 s, 58 °C for 30 s, 72 °C for 20 s, with a final extension at 72 °C for 7 min.

    Article Title: Cell-specific exon methylation and CTCF binding in neurons regulate calcium ion channel splicing and function
    Article Snippet: .. RNA extraction, RT-PCR and qPCR RNA extraction from F11 cells was performed using TRIzol (Invitrogen Cat# 15596018) and from dissociated DRG cells using TRIzol LS Reagent (Invitrogen Cat# 10296010) according to the manufacturer’s instructions. .. 1–2 μg of RNA was immediately reverse transcribed to cDNA using the SuperScript III First-Strand Synthesis System with Poli-dT primers (Invitrogen Cat# 18080051).

    Article Title: An Infectious cDNA Clone of SARS-CoV-2
    Article Snippet: .. RNA Extraction, RT-PCR and Sanger Sequencing 250 μL of culture fluids were mixed with three volumes of TRIzol LS Reagent (Thermo Fisher Scientific). ..

    Sequencing:

    Article Title: An Infectious cDNA Clone of SARS-CoV-2
    Article Snippet: .. RNA Extraction, RT-PCR and Sanger Sequencing 250 μL of culture fluids were mixed with three volumes of TRIzol LS Reagent (Thermo Fisher Scientific). ..

    Plasmid Preparation:

    Article Title: In vitro inhibition of CSFV replication by retroviral vector-mediated RNA interference
    Article Snippet: .. 2.5.2 Construction of 5′-UTR plasmid Total RNA of CSFV-infected cells was extracted with Trizol LS Reagent (Invitrogen, Carlsbad, CA, USA) and then applied as a template for synthesis of the first cDNA strand of the 5′-UTR by RT-PCR using AMV reverse transcriptase (Promega, Madison, WI, USA) and 5′-UTR RP267 primer, according to the manufacturer's instructions. .. PCR was then used to amplify the 5′-UTR with FP147 and RP267 primers under conditions of 94 °C for 2 min, then 35 cycles of 94 °C for 30 s, 58 °C for 30 s, 72 °C for 20 s, with a final extension at 72 °C for 7 min.

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    Thermo Fisher trizol ls
    (a) Western blot analysis of CTB-, AV- and ST-bound MSC EVs. MSC CM was incubated with CTB, AV or ST followed by incubation with Dynabeads conjugated with Streptavidin. The beads were immobilised with a magnet, washed, denatured and resolved onto polyacrylamide gels before electroblotting onto a nitrocellulose membrane. The membrane was probed with a primary antibody followed by horseradish peroxidase-coupled secondary antibodies against the primary antibody. The blot was then incubated with a chemiluminescent HRP substrate to detect bound primary antibody. (b) 10 µg MSC EV was extracted sequentially with biotinylated CTB and then biotinylated AV or vice versa. After each extraction, the ligand-bound vesicles were removed with Dynabeads ® MyOne Streptavidin T1 and assayed for CD81 by ELISA. The relative level of CD81 in CTB-vesicles before and after extraction with AV, and that in AV-vesicles before and after extraction with CTB were normalized to that in AV-vesicles before CTB extraction. (c) <t>RNA</t> analysis of CTB-, AV- and ST-EVs. CTB-, AV- or ST-binding EVs were isolated as described above and extracted for RNA using <t>Trizol.</t> The pellet in each of extracts was re-suspended in 50 µL of RNase-free water. 10 µL of each RNA solution was resolved on a 15% Novex Tris-borate-EDTA(TBE)-urea gel before staining with ethidium bromide.
    Trizol Ls, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 464 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trizol ls/product/Thermo Fisher
    Average 99 stars, based on 464 article reviews
    Price from $9.99 to $1999.99
    trizol ls - by Bioz Stars, 2020-05
    99/100 stars
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    (a) Western blot analysis of CTB-, AV- and ST-bound MSC EVs. MSC CM was incubated with CTB, AV or ST followed by incubation with Dynabeads conjugated with Streptavidin. The beads were immobilised with a magnet, washed, denatured and resolved onto polyacrylamide gels before electroblotting onto a nitrocellulose membrane. The membrane was probed with a primary antibody followed by horseradish peroxidase-coupled secondary antibodies against the primary antibody. The blot was then incubated with a chemiluminescent HRP substrate to detect bound primary antibody. (b) 10 µg MSC EV was extracted sequentially with biotinylated CTB and then biotinylated AV or vice versa. After each extraction, the ligand-bound vesicles were removed with Dynabeads ® MyOne Streptavidin T1 and assayed for CD81 by ELISA. The relative level of CD81 in CTB-vesicles before and after extraction with AV, and that in AV-vesicles before and after extraction with CTB were normalized to that in AV-vesicles before CTB extraction. (c) RNA analysis of CTB-, AV- and ST-EVs. CTB-, AV- or ST-binding EVs were isolated as described above and extracted for RNA using Trizol. The pellet in each of extracts was re-suspended in 50 µL of RNase-free water. 10 µL of each RNA solution was resolved on a 15% Novex Tris-borate-EDTA(TBE)-urea gel before staining with ethidium bromide.

    Journal: Journal of Extracellular Vesicles

    Article Title: MSC secretes at least 3 EV types each with a unique permutation of membrane lipid, protein and RNA

    doi: 10.3402/jev.v5.29828

    Figure Lengend Snippet: (a) Western blot analysis of CTB-, AV- and ST-bound MSC EVs. MSC CM was incubated with CTB, AV or ST followed by incubation with Dynabeads conjugated with Streptavidin. The beads were immobilised with a magnet, washed, denatured and resolved onto polyacrylamide gels before electroblotting onto a nitrocellulose membrane. The membrane was probed with a primary antibody followed by horseradish peroxidase-coupled secondary antibodies against the primary antibody. The blot was then incubated with a chemiluminescent HRP substrate to detect bound primary antibody. (b) 10 µg MSC EV was extracted sequentially with biotinylated CTB and then biotinylated AV or vice versa. After each extraction, the ligand-bound vesicles were removed with Dynabeads ® MyOne Streptavidin T1 and assayed for CD81 by ELISA. The relative level of CD81 in CTB-vesicles before and after extraction with AV, and that in AV-vesicles before and after extraction with CTB were normalized to that in AV-vesicles before CTB extraction. (c) RNA analysis of CTB-, AV- and ST-EVs. CTB-, AV- or ST-binding EVs were isolated as described above and extracted for RNA using Trizol. The pellet in each of extracts was re-suspended in 50 µL of RNase-free water. 10 µL of each RNA solution was resolved on a 15% Novex Tris-borate-EDTA(TBE)-urea gel before staining with ethidium bromide.

    Article Snippet: The isolated EVs were resuspended in 100 µL of PBS and extracted for RNA using 3 volumes of Trizol LS (Thermo Fisher Scientific, Waltham, MA) according to the manufacturer's protocol.

    Techniques: Western Blot, CtB Assay, Incubation, Enzyme-linked Immunosorbent Assay, Binding Assay, Isolation, Staining

    Characterization of HBV DNA and RNA in sera of CHB patients. (A and B) Analyses of serum viral DNA from CHB patients by Southern blotting. Viral DNA was extracted from serum samples obtained from forty-five chronic hepatitis B patients (20% of input sample used for protein A/G agarose beads pulldown) and subjected to Southern blot analysis. Alternatively, these samples were first incubated with protein A/G agarose beads, and then viral DNA in the pulldown mixtures was analyzed by Southern blotting. Serum samples selected for further examining are marked with arrows, and samples with SS DNA detection are labeled with asterisks. (C) Protein A/G agarose bead pulldown of viral particles. Sera (25 μl each) from CHB patients 37, 38, 14, and 35 (M1, mixture one) or from patients 17, 21, 42, and 44 (M2, mixture two) were pooled and incubated with protein A/G agarose beads. Viral DNA in input sera, protein A/G bead pulldown mixtures (beads), and the remaining supernatants (sup.) were extracted and subjected to Southern blot analysis. (D) Northern blot detection of serum viral RNA from patients 37, 38, 14, 35, 17, 21, 42, and 44. Total RNA were extracted from serum samples by TRIzol reagent and treated with DNase I before Northern blot analysis. (E to G) Southern blot analyses of viral DNA from selected samples. Viral DNA was separated by electrophoresis through TAE or alkaline agarose gels, followed by Southern blot detection with the indicated riboprobes.

    Journal: Journal of Virology

    Article Title: Extracellular Hepatitis B Virus RNAs Are Heterogeneous in Length and Circulate as Capsid-Antibody Complexes in Addition to Virions in Chronic Hepatitis B Patients

    doi: 10.1128/JVI.00798-18

    Figure Lengend Snippet: Characterization of HBV DNA and RNA in sera of CHB patients. (A and B) Analyses of serum viral DNA from CHB patients by Southern blotting. Viral DNA was extracted from serum samples obtained from forty-five chronic hepatitis B patients (20% of input sample used for protein A/G agarose beads pulldown) and subjected to Southern blot analysis. Alternatively, these samples were first incubated with protein A/G agarose beads, and then viral DNA in the pulldown mixtures was analyzed by Southern blotting. Serum samples selected for further examining are marked with arrows, and samples with SS DNA detection are labeled with asterisks. (C) Protein A/G agarose bead pulldown of viral particles. Sera (25 μl each) from CHB patients 37, 38, 14, and 35 (M1, mixture one) or from patients 17, 21, 42, and 44 (M2, mixture two) were pooled and incubated with protein A/G agarose beads. Viral DNA in input sera, protein A/G bead pulldown mixtures (beads), and the remaining supernatants (sup.) were extracted and subjected to Southern blot analysis. (D) Northern blot detection of serum viral RNA from patients 37, 38, 14, 35, 17, 21, 42, and 44. Total RNA were extracted from serum samples by TRIzol reagent and treated with DNase I before Northern blot analysis. (E to G) Southern blot analyses of viral DNA from selected samples. Viral DNA was separated by electrophoresis through TAE or alkaline agarose gels, followed by Southern blot detection with the indicated riboprobes.

    Article Snippet: In addition to the SDS-proteinase K method, viral RNA was also extracted with TRIzol LS reagent according to the manufacturer’s instructions (Thermo Fisher Scientific).

    Techniques: Southern Blot, Incubation, Labeling, Northern Blot, Electrophoresis

    ). (B) Identification of 3′ ends of extracellular HBV RNAs. 3′ Ends of extracellular HBV RNAs were identified by the 3′ RACE method using different HBV-specific anchor primers (the same 5′ primers used for generating templates for producing riboprobes used in panel A, lower). Identified 3′ ends were numbered as described above, and numbers in parentheses indicate the amount of clones with the same 3′ ends. The asterisk indicates unknown nucleic acid copurified with intracellular capsid-associated viral RNA by TRIzol reagent. FL, full-length; Cap, 5′ cap of pregenomic RNA; pA, the polyadenylation site; An, poly(A) tail.

    Journal: Journal of Virology

    Article Title: Extracellular Hepatitis B Virus RNAs Are Heterogeneous in Length and Circulate as Capsid-Antibody Complexes in Addition to Virions in Chronic Hepatitis B Patients

    doi: 10.1128/JVI.00798-18

    Figure Lengend Snippet: ). (B) Identification of 3′ ends of extracellular HBV RNAs. 3′ Ends of extracellular HBV RNAs were identified by the 3′ RACE method using different HBV-specific anchor primers (the same 5′ primers used for generating templates for producing riboprobes used in panel A, lower). Identified 3′ ends were numbered as described above, and numbers in parentheses indicate the amount of clones with the same 3′ ends. The asterisk indicates unknown nucleic acid copurified with intracellular capsid-associated viral RNA by TRIzol reagent. FL, full-length; Cap, 5′ cap of pregenomic RNA; pA, the polyadenylation site; An, poly(A) tail.

    Article Snippet: In addition to the SDS-proteinase K method, viral RNA was also extracted with TRIzol LS reagent according to the manufacturer’s instructions (Thermo Fisher Scientific).

    Techniques: Clone Assay

    Characterization of HBV DNA and RNA in sera of CHB patients. (A and B) Analyses of serum viral DNA from CHB patients by Southern blotting. Viral DNA was extracted from serum samples obtained from forty-five chronic hepatitis B patients (20% of input sample used for protein A/G agarose beads pulldown) and subjected to Southern blot analysis. Alternatively, these samples were first incubated with protein A/G agarose beads, and then viral DNA in the pulldown mixtures was analyzed by Southern blotting. Serum samples selected for further examining are marked with arrows, and samples with SS DNA detection are labeled with asterisks. (C) Protein A/G agarose bead pulldown of viral particles. Sera (25 μl each) from CHB patients 37, 38, 14, and 35 (M1, mixture one) or from patients 17, 21, 42, and 44 (M2, mixture two) were pooled and incubated with protein A/G agarose beads. Viral DNA in input sera, protein A/G bead pulldown mixtures (beads), and the remaining supernatants (sup.) were extracted and subjected to Southern blot analysis. (D) Northern blot detection of serum viral RNA from patients 37, 38, 14, 35, 17, 21, 42, and 44. Total RNA were extracted from serum samples by TRIzol reagent and treated with DNase I before Northern blot analysis. (E to G) Southern blot analyses of viral DNA from selected samples. Viral DNA was separated by electrophoresis through TAE or alkaline agarose gels, followed by Southern blot detection with the indicated riboprobes.

    Journal: Journal of Virology

    Article Title: Extracellular Hepatitis B Virus RNAs Are Heterogeneous in Length and Circulate as Capsid-Antibody Complexes in Addition to Virions in Chronic Hepatitis B Patients

    doi: 10.1128/JVI.00798-18

    Figure Lengend Snippet: Characterization of HBV DNA and RNA in sera of CHB patients. (A and B) Analyses of serum viral DNA from CHB patients by Southern blotting. Viral DNA was extracted from serum samples obtained from forty-five chronic hepatitis B patients (20% of input sample used for protein A/G agarose beads pulldown) and subjected to Southern blot analysis. Alternatively, these samples were first incubated with protein A/G agarose beads, and then viral DNA in the pulldown mixtures was analyzed by Southern blotting. Serum samples selected for further examining are marked with arrows, and samples with SS DNA detection are labeled with asterisks. (C) Protein A/G agarose bead pulldown of viral particles. Sera (25 μl each) from CHB patients 37, 38, 14, and 35 (M1, mixture one) or from patients 17, 21, 42, and 44 (M2, mixture two) were pooled and incubated with protein A/G agarose beads. Viral DNA in input sera, protein A/G bead pulldown mixtures (beads), and the remaining supernatants (sup.) were extracted and subjected to Southern blot analysis. (D) Northern blot detection of serum viral RNA from patients 37, 38, 14, 35, 17, 21, 42, and 44. Total RNA were extracted from serum samples by TRIzol reagent and treated with DNase I before Northern blot analysis. (E to G) Southern blot analyses of viral DNA from selected samples. Viral DNA was separated by electrophoresis through TAE or alkaline agarose gels, followed by Southern blot detection with the indicated riboprobes.

    Article Snippet: In addition to the SDS-proteinase K method, viral RNA was also extracted with TRIzol LS reagent according to the manufacturer’s instructions (Thermo Fisher Scientific).

    Techniques: Southern Blot, Incubation, Labeling, Northern Blot, Electrophoresis

    Mapping and identifying 3′ ends of extracellular HBV RNAs. (A) Northern blot detection of extracellular HBV RNAs with various riboprobes. Viral RNA from cytoplasmic (C) nucleocapsids (lanes 2, 5, 8, 11, 14, and 17) or culture supernatant (S) (lanes 3, 6, 9, 12, 15, and 18) of HepAD38 cells was extracted with TRIzol reagent and treated with DNase I before Northern blot analysis with plus-strand-specific riboprobes spanning the HBV genome as indicated. pgRNA was used as a reference, and map coordinates were numbered according to the sequence of the HBV genome (genotype D, accession number AJ344117.1 ). (B) Identification of 3′ ends of extracellular HBV RNAs. 3′ Ends of extracellular HBV RNAs were identified by the 3′ RACE method using different HBV-specific anchor primers (the same 5′ primers used for generating templates for producing riboprobes used in panel A, lower). Identified 3′ ends were numbered as described above, and numbers in parentheses indicate the amount of clones with the same 3′ ends. The asterisk indicates unknown nucleic acid copurified with intracellular capsid-associated viral RNA by TRIzol reagent. FL, full-length; Cap, 5′ cap of pregenomic RNA; pA, the polyadenylation site; An, poly(A) tail.

    Journal: Journal of Virology

    Article Title: Extracellular Hepatitis B Virus RNAs Are Heterogeneous in Length and Circulate as Capsid-Antibody Complexes in Addition to Virions in Chronic Hepatitis B Patients

    doi: 10.1128/JVI.00798-18

    Figure Lengend Snippet: Mapping and identifying 3′ ends of extracellular HBV RNAs. (A) Northern blot detection of extracellular HBV RNAs with various riboprobes. Viral RNA from cytoplasmic (C) nucleocapsids (lanes 2, 5, 8, 11, 14, and 17) or culture supernatant (S) (lanes 3, 6, 9, 12, 15, and 18) of HepAD38 cells was extracted with TRIzol reagent and treated with DNase I before Northern blot analysis with plus-strand-specific riboprobes spanning the HBV genome as indicated. pgRNA was used as a reference, and map coordinates were numbered according to the sequence of the HBV genome (genotype D, accession number AJ344117.1 ). (B) Identification of 3′ ends of extracellular HBV RNAs. 3′ Ends of extracellular HBV RNAs were identified by the 3′ RACE method using different HBV-specific anchor primers (the same 5′ primers used for generating templates for producing riboprobes used in panel A, lower). Identified 3′ ends were numbered as described above, and numbers in parentheses indicate the amount of clones with the same 3′ ends. The asterisk indicates unknown nucleic acid copurified with intracellular capsid-associated viral RNA by TRIzol reagent. FL, full-length; Cap, 5′ cap of pregenomic RNA; pA, the polyadenylation site; An, poly(A) tail.

    Article Snippet: In addition to the SDS-proteinase K method, viral RNA was also extracted with TRIzol LS reagent according to the manufacturer’s instructions (Thermo Fisher Scientific).

    Techniques: Northern Blot, Sequencing, Clone Assay