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Thermo Fisher trizol
Myr-Akt mice demonstrate increased F4/80+ macrophages in the spleens and a M2 macrophage bias in the lungs. 6 – 8 week old Myr-Akt or WT mice were sacrificed and (a, top ) spleens and (a, bottom ) lungs were harvested, paraffin embedded, and subjected to F4/80 immunohistochemical staining for macrophages. Ten pictures per section were imaged using 4X and 20X objectives (40× and 200× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) and expressed as mean ± SEM. (c) BMMs were generated from Myr-Akt or WT littermate mice with recombinant murine (rh)CSF1 (20 ng/ml). Cells were removed using Accutase one and two days after seeding the progenitor cells (day 0). Cell surface antigen expression of F4/80 was assessed by flow cytometry. Data presented are the average geometric mean for F4/80 expression ± SEM. (d) Total <t>RNA</t> was extracted from lung and spleen using <t>TRIzol</t> and cDNA synthesized from 1 μg RNA for quantitative real-time (qRT)-PCR using primers specific for mouse IL 12, iNos , IL 10, or Ym 1. Data are expressed as relative copy number (RCN) (d, left ) IL 12 and iNos mRNA, or (d, right ) IL 10 mRNA and Ym 1 mRNA expression over the mean of endogenous control RNAs ( Cap1 , Rpl 4 and Gapdh) . Data represents the mean RCN ± SEM. 5 pairs of age-matched littermate mice were used for each experiment.
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Images

1) Product Images from "Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy"

Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

Journal: Advances in bioscience and biotechnology (Print)

doi: 10.4236/abb.2019.1010027

Myr-Akt mice demonstrate increased F4/80+ macrophages in the spleens and a M2 macrophage bias in the lungs. 6 – 8 week old Myr-Akt or WT mice were sacrificed and (a, top ) spleens and (a, bottom ) lungs were harvested, paraffin embedded, and subjected to F4/80 immunohistochemical staining for macrophages. Ten pictures per section were imaged using 4X and 20X objectives (40× and 200× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) and expressed as mean ± SEM. (c) BMMs were generated from Myr-Akt or WT littermate mice with recombinant murine (rh)CSF1 (20 ng/ml). Cells were removed using Accutase one and two days after seeding the progenitor cells (day 0). Cell surface antigen expression of F4/80 was assessed by flow cytometry. Data presented are the average geometric mean for F4/80 expression ± SEM. (d) Total RNA was extracted from lung and spleen using TRIzol and cDNA synthesized from 1 μg RNA for quantitative real-time (qRT)-PCR using primers specific for mouse IL 12, iNos , IL 10, or Ym 1. Data are expressed as relative copy number (RCN) (d, left ) IL 12 and iNos mRNA, or (d, right ) IL 10 mRNA and Ym 1 mRNA expression over the mean of endogenous control RNAs ( Cap1 , Rpl 4 and Gapdh) . Data represents the mean RCN ± SEM. 5 pairs of age-matched littermate mice were used for each experiment.
Figure Legend Snippet: Myr-Akt mice demonstrate increased F4/80+ macrophages in the spleens and a M2 macrophage bias in the lungs. 6 – 8 week old Myr-Akt or WT mice were sacrificed and (a, top ) spleens and (a, bottom ) lungs were harvested, paraffin embedded, and subjected to F4/80 immunohistochemical staining for macrophages. Ten pictures per section were imaged using 4X and 20X objectives (40× and 200× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) and expressed as mean ± SEM. (c) BMMs were generated from Myr-Akt or WT littermate mice with recombinant murine (rh)CSF1 (20 ng/ml). Cells were removed using Accutase one and two days after seeding the progenitor cells (day 0). Cell surface antigen expression of F4/80 was assessed by flow cytometry. Data presented are the average geometric mean for F4/80 expression ± SEM. (d) Total RNA was extracted from lung and spleen using TRIzol and cDNA synthesized from 1 μg RNA for quantitative real-time (qRT)-PCR using primers specific for mouse IL 12, iNos , IL 10, or Ym 1. Data are expressed as relative copy number (RCN) (d, left ) IL 12 and iNos mRNA, or (d, right ) IL 10 mRNA and Ym 1 mRNA expression over the mean of endogenous control RNAs ( Cap1 , Rpl 4 and Gapdh) . Data represents the mean RCN ± SEM. 5 pairs of age-matched littermate mice were used for each experiment.

Techniques Used: Mouse Assay, Immunohistochemistry, Staining, Software, Generated, Recombinant, Expressing, Flow Cytometry, Cytometry, Synthesized, Quantitative RT-PCR

Bleomycin induces collagen deposition in the lungs and reduces survival in Myr-Akt mice. Myr-Akt and wild type (WT) littermate mice were treated twice a week with bleomycin (0.035 U/g) or PBS (vehicle) for 4 weeks, as described in Material and Methods. (a) Mouse survival was recorded and plotted using a Kaplan-Meier curve. Myr-Akt mice had worse survival than WT mice in response to bleomycin treatment. N = 7 mice for each WT and Myr-Akt mouse group. (b) All surviving mice were sacrificed at day 33 and the lungs formalin-fixed, sectioned, and stained with Masson’s Trichrome. Five images per slide were captured. Red staining represents keratin, light red and pink staining represents cytoplasm, dark brown represents cell nuclei, and blue staining represents collagen. The images are representative images from 3 mice per group using a 20X objective (200× magnification). (c) The images were quantified blindly by histogram analysis using Adobe Photoshop CS and the percent of blue staining per high-power field (HPF) from ten images and expressed as mean ± SEM. (d) Lungs from WT and Myr-Akt mice subjected to PBS or bleomycin stained with H E, Trichrome, and F4/80 were evaluated by a board-certified pathologist for inflammation score (0 to 5 point scale). (e) Total RNA was extracted from homogenized lung tissue using TRIzol. cDNA was synthesized and qRT-PCR performed for mouse collagen IA and IIIB. Data are expressed as mean relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM and N = 4 mice per group.
Figure Legend Snippet: Bleomycin induces collagen deposition in the lungs and reduces survival in Myr-Akt mice. Myr-Akt and wild type (WT) littermate mice were treated twice a week with bleomycin (0.035 U/g) or PBS (vehicle) for 4 weeks, as described in Material and Methods. (a) Mouse survival was recorded and plotted using a Kaplan-Meier curve. Myr-Akt mice had worse survival than WT mice in response to bleomycin treatment. N = 7 mice for each WT and Myr-Akt mouse group. (b) All surviving mice were sacrificed at day 33 and the lungs formalin-fixed, sectioned, and stained with Masson’s Trichrome. Five images per slide were captured. Red staining represents keratin, light red and pink staining represents cytoplasm, dark brown represents cell nuclei, and blue staining represents collagen. The images are representative images from 3 mice per group using a 20X objective (200× magnification). (c) The images were quantified blindly by histogram analysis using Adobe Photoshop CS and the percent of blue staining per high-power field (HPF) from ten images and expressed as mean ± SEM. (d) Lungs from WT and Myr-Akt mice subjected to PBS or bleomycin stained with H E, Trichrome, and F4/80 were evaluated by a board-certified pathologist for inflammation score (0 to 5 point scale). (e) Total RNA was extracted from homogenized lung tissue using TRIzol. cDNA was synthesized and qRT-PCR performed for mouse collagen IA and IIIB. Data are expressed as mean relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM and N = 4 mice per group.

Techniques Used: Mouse Assay, Staining, Synthesized, Quantitative RT-PCR, IA, Expressing

Reduced autophagy in the lungs of Myr-Akt mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to LC3B immunohistochemical staining. Ten pictures per section were imaged using 10X and 40X objectives (100× and 400× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) from at least five images per section and expressed as mean ± SEM. (c) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse Beclin-1 , Lc 3 a , and Lc 3 b mRNAs. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (d) Total protein were isolated and equal amounts of protein separated on a 4% – 12% SDS-PAGE gel and immunoblotted for BECLIN-1, LC3A/B-I and LC3A/B-II, and β -ACTIN. (e) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse p 62/ SQSTM1 mRNA. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (f) Immunoblot for phosphor-mTOR (ser2448) and mTOR (Cell Signaling Technology). Bands were quantified using densitometry by comparing band density over β -ACTIN or total mTOR in each respective lane. Data are expressed as the mean densitometry value ± SEM. 4 pairs of age-matched littermate mice were used for western analysis and 5 pairs of age-matched littermate mice were used for gene expression.
Figure Legend Snippet: Reduced autophagy in the lungs of Myr-Akt mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to LC3B immunohistochemical staining. Ten pictures per section were imaged using 10X and 40X objectives (100× and 400× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) from at least five images per section and expressed as mean ± SEM. (c) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse Beclin-1 , Lc 3 a , and Lc 3 b mRNAs. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (d) Total protein were isolated and equal amounts of protein separated on a 4% – 12% SDS-PAGE gel and immunoblotted for BECLIN-1, LC3A/B-I and LC3A/B-II, and β -ACTIN. (e) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse p 62/ SQSTM1 mRNA. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (f) Immunoblot for phosphor-mTOR (ser2448) and mTOR (Cell Signaling Technology). Bands were quantified using densitometry by comparing band density over β -ACTIN or total mTOR in each respective lane. Data are expressed as the mean densitometry value ± SEM. 4 pairs of age-matched littermate mice were used for western analysis and 5 pairs of age-matched littermate mice were used for gene expression.

Techniques Used: Mouse Assay, Immunohistochemistry, Staining, Software, Synthesized, Quantitative RT-PCR, Expressing, Isolation, SDS Page, Western Blot

2) Product Images from "Stem cell transplantation impairs dendritic cell trafficking and herpesvirus immunity"

Article Title: Stem cell transplantation impairs dendritic cell trafficking and herpesvirus immunity

Journal: JCI Insight

doi: 10.1172/jci.insight.130210

APCs from lungs of BMT mice are potent in priming both Th1 and Th17 responses in vitro. ( A ) Single-cell suspensions were prepared by collagenase digestion of whole lungs of non-BMT ( n = 5) or BMT ( n = 5) mice at 7 dpi with MHV-68. Cells were then stimulated with PMA and ionomycin for 4 hours before antibody staining. Percent of CD4 + T cells (i.e., CD45 + CD90.2 + CD3 + CD4 + ) that express IFN-γ (Th1 cells), and percent of CD4 + T cells that express IL-17A (Th17 cells) were determined by flow cytometry. ( B and C ) APCs enriched by CD11c + microbeads and pooled from lung Single-cell suspensions of 5 BMT or non-BMT mice at 3 dpi were cocultured with pooled CD4 + T cells from 10 BMT mouse lungs at 10 dpi at a 1:10 ratio in the presence of 0.125 MOI of MHV-68 ( n = 5 each). ( B ) The concentration of IFN-γ or IL-17A in the supernatant of coculture at 4 days was determined by ELISA (mean ± SEM, n = 5). ( C ) Cocultured cells were pelleted at 4 days after coculture, and total RNA was isolated by TRIzol. The expression of a pro-Th1 cytokine ( IL12A ) or pro-Th17 cytokines ( IL6 and IL23A ) were determined by real-time RT-PCR (mean ± SEM, n = 5). Symbols represent individual data points from unique mice. * P
Figure Legend Snippet: APCs from lungs of BMT mice are potent in priming both Th1 and Th17 responses in vitro. ( A ) Single-cell suspensions were prepared by collagenase digestion of whole lungs of non-BMT ( n = 5) or BMT ( n = 5) mice at 7 dpi with MHV-68. Cells were then stimulated with PMA and ionomycin for 4 hours before antibody staining. Percent of CD4 + T cells (i.e., CD45 + CD90.2 + CD3 + CD4 + ) that express IFN-γ (Th1 cells), and percent of CD4 + T cells that express IL-17A (Th17 cells) were determined by flow cytometry. ( B and C ) APCs enriched by CD11c + microbeads and pooled from lung Single-cell suspensions of 5 BMT or non-BMT mice at 3 dpi were cocultured with pooled CD4 + T cells from 10 BMT mouse lungs at 10 dpi at a 1:10 ratio in the presence of 0.125 MOI of MHV-68 ( n = 5 each). ( B ) The concentration of IFN-γ or IL-17A in the supernatant of coculture at 4 days was determined by ELISA (mean ± SEM, n = 5). ( C ) Cocultured cells were pelleted at 4 days after coculture, and total RNA was isolated by TRIzol. The expression of a pro-Th1 cytokine ( IL12A ) or pro-Th17 cytokines ( IL6 and IL23A ) were determined by real-time RT-PCR (mean ± SEM, n = 5). Symbols represent individual data points from unique mice. * P

Techniques Used: Mouse Assay, In Vitro, Staining, Flow Cytometry, Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Quantitative RT-PCR

3) Product Images from "External quality assessment for PML‐RARα detection in acute promyelocytic leukemia: Findings and summary. External quality assessment for PML‐RARα detection in acute promyelocytic leukemia: Findings and summary"

Article Title: External quality assessment for PML‐RARα detection in acute promyelocytic leukemia: Findings and summary. External quality assessment for PML‐RARα detection in acute promyelocytic leukemia: Findings and summary

Journal: Journal of Clinical Laboratory Analysis

doi: 10.1002/jcla.22894

Detection performance of mock leukocyte samples for EQA panels. A, RNA yields extracted by different methods. Error bars represent standard deviation. B, RNA yields extracted by TRIzol between EQA samples. C, Control gene copy number of each mock leukocyte sample. D, Differences in the slope of RT‐qPCR standard curve between different laboratory groups. 1, correct detection group; 2, quantitative incorrect group; 3, only qualitative incorrect group. Error bars represent standard deviation. ** P
Figure Legend Snippet: Detection performance of mock leukocyte samples for EQA panels. A, RNA yields extracted by different methods. Error bars represent standard deviation. B, RNA yields extracted by TRIzol between EQA samples. C, Control gene copy number of each mock leukocyte sample. D, Differences in the slope of RT‐qPCR standard curve between different laboratory groups. 1, correct detection group; 2, quantitative incorrect group; 3, only qualitative incorrect group. Error bars represent standard deviation. ** P

Techniques Used: Standard Deviation, Quantitative RT-PCR

4) Product Images from "Obesity induces pro-inflammatory B cells and impairs B cell function in old mice"

Article Title: Obesity induces pro-inflammatory B cells and impairs B cell function in old mice

Journal: Mechanisms of ageing and development

doi: 10.1016/j.mad.2017.01.004

The adipocytes secrete chemokines which may recruit B cells to the VAT. A. Adipocytes were isolated from the VAT, sonicated for cell disruption in the presence of TRIzol to separate the soluble fraction (used for RNA isolation) from lipids and cell debris. Results show qPCR values (2 − ΔΔ Ct ) of CXCL10, CCL2, CCL5 RNA expression. Mean comparisons between groups were performed by Student's t -test (two-tailed). B. VAT B cells were isolated from the SVF and resuspended in TRIzol. Results show qPCR values (2 − ΔΔ Ct ) of CXCR2, CCR2, CCR3 RNA expression. Results in B are from those 3 mice among the 6 in A from which we had enough B cells. Mean comparisons between groups were performed by Student's t-test (two-tailed). *p
Figure Legend Snippet: The adipocytes secrete chemokines which may recruit B cells to the VAT. A. Adipocytes were isolated from the VAT, sonicated for cell disruption in the presence of TRIzol to separate the soluble fraction (used for RNA isolation) from lipids and cell debris. Results show qPCR values (2 − ΔΔ Ct ) of CXCL10, CCL2, CCL5 RNA expression. Mean comparisons between groups were performed by Student's t -test (two-tailed). B. VAT B cells were isolated from the SVF and resuspended in TRIzol. Results show qPCR values (2 − ΔΔ Ct ) of CXCR2, CCR2, CCR3 RNA expression. Results in B are from those 3 mice among the 6 in A from which we had enough B cells. Mean comparisons between groups were performed by Student's t-test (two-tailed). *p

Techniques Used: Isolation, Sonication, Real-time Polymerase Chain Reaction, RNA Expression, Two Tailed Test, Mouse Assay

5) Product Images from "Competitive Inhibition of Amino Acid Uptake Suppresses Chlamydial Growth: Involvement of the Chlamydial Amino Acid Transporter BrnQ ▿Competitive Inhibition of Amino Acid Uptake Suppresses Chlamydial Growth: Involvement of the Chlamydial Amino Acid Transporter BrnQ ▿ †"

Article Title: Competitive Inhibition of Amino Acid Uptake Suppresses Chlamydial Growth: Involvement of the Chlamydial Amino Acid Transporter BrnQ ▿Competitive Inhibition of Amino Acid Uptake Suppresses Chlamydial Growth: Involvement of the Chlamydial Amino Acid Transporter BrnQ ▿ †

Journal:

doi: 10.1128/JB.01240-07

CT554 is expressed throughout the entire chlamydial developmental cycle. HeLa cells were infected using an MOI of 1, and total RNA was prepared at 2, 8, 16, 24, and 44 h p.i. using Trizol for analysis by quantitative real-time RT-PCR. The expression of
Figure Legend Snippet: CT554 is expressed throughout the entire chlamydial developmental cycle. HeLa cells were infected using an MOI of 1, and total RNA was prepared at 2, 8, 16, 24, and 44 h p.i. using Trizol for analysis by quantitative real-time RT-PCR. The expression of

Techniques Used: Infection, Quantitative RT-PCR, Expressing

Related Articles

Clone Assay:

Article Title: The emergent landscape of the mouse gut endoderm at single-cell resolution
Article Snippet: Paragraph title: Amplification and cloning of antisense riboprobes ... Total RNA was isolated from whole 13ss stage wild-type embryos using Trizol (Invitrogen).

Amplification:

Article Title: The emergent landscape of the mouse gut endoderm at single-cell resolution
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Mass Spectrometry:

Article Title: In vivo BiFC analysis of Y14 and NXF1 mRNA export complexes: preferential localization within and around SC35 domains
Article Snippet: RNA purification with TRIzol After the immunoprecipitation, 1 ml TRIzol (Invitrogen) was added, mixed, and incubated at RT for 5 min. 200 μl chloroform was added, mixed, and incubated at RT for 15 min. .. Radioactivity was exposed on BioMax MS film (Kodak).

Construct:

Article Title: External quality assessment for PML‐RARα detection in acute promyelocytic leukemia: Findings and summary. External quality assessment for PML‐RARα detection in acute promyelocytic leukemia: Findings and summary
Article Snippet: We constructed PML‐RARα FG EQA panel which consisted of limited positive and negative samples with different FGCN /CGCN ratio and MRD value (Table ). .. Total RNA was extracted by TRIzol reagent and spin column, quantified using NanoDrop 2000c (Thermo Fisher).

SYBR Green Assay:

Article Title: Cell membrane coating for reducing nanoparticle-induced inflammatory responses to scaffold constructs
Article Snippet: Fresh cells isolated from scaffolds were stored in TRIzol (Life Technologies), and RNA from cells was isolated using TRIzol reagent and TURBO DNA-free Kit (Invitrogen) according to manufacturer instructions. .. The concentration of RNA was quantified using the NanoDrop 1000 Spectro-photomer (Thermo Scientific). cDNA was prepared according to the protocol provided by the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). qPCR was carried out using PowerUp SYBR Green Master Mix (Applied Biosystems) on QuantStudio 6 Flex Real-Time PCR System (Applied Biosystems).

Article Title: HHIP overexpression inhibits the proliferation, migration and invasion of non-small cell lung cancer
Article Snippet: .. Experimental materials and reagents Human normal lung cell line (BEAS-2B) and NSCLC cell lines (H1975, H358, H226 and HCC827) were purchased from ATCC (USA); RPMI-1640 medium, fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco (USA); Trizol and LipofectamineTM 2000 were purchased from ThermoFisher (USA); pEGFP overexpression plasmid was purchased from Shanghai Beinuo Biotechnology Co., Ltd.; CCK-8 kit was purchased from Shanghai Beyotime Biotechnology Co., Ltd.; cDNA PrimeScript RT kit and SYBR-Green PCR Master Mix kit were purchased from Dalian TaKaRa Co., Ltd.; Transwell chamber was purchased from Milipore (USA); artificial Matrigel was purchased from BD Biosciences (USA). .. ABI 7500 fluorescence quantitative PCR was purchased from Applied Biosystems (USA).

Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy
Article Snippet: RNA Extraction and Quantitative Real-Time PCR Total RNA was extracted using TRIzol (Thermofisher Scientific) and 1 μg total RNA was used to synthesize cDNA with SuperScript III Reverse Transcriptase (Thermofisher Scientific). .. Quantitative real-time (qRT)-PCR was performed in duplicate using SYBR Green Master Mix (Applied BioSystems) and an ABI PRIZM 7700 with Sequence Detector v1.7 (Applied Biosystems).

Microarray:

Article Title: Oroxin B Induces Apoptosis by Down-Regulating MicroRNA-221 Resulting in the Inactivation of the PTEN/PI3K/AKT Pathway in Liver Cancer
Article Snippet: .. Microarray Analysis Total RNA was extracted from liver tissue, using TRIzol (Ambion, Carlsbad, CA, USA) according to the manufacturer’s instructions. .. After phosphatase treatment, 100 ng total RNA for each sample was incubated for 30 min at 37 °C.

Incubation:

Article Title: In vivo BiFC analysis of Y14 and NXF1 mRNA export complexes: preferential localization within and around SC35 domains
Article Snippet: .. RNA purification with TRIzol After the immunoprecipitation, 1 ml TRIzol (Invitrogen) was added, mixed, and incubated at RT for 5 min. 200 μl chloroform was added, mixed, and incubated at RT for 15 min. ..

Article Title: Obesity induces pro-inflammatory B cells and impairs B cell function in old mice
Article Snippet: B cells were isolated from the spleens or from the SVF of young and old mice after a 20 min incubation at 4°C with anti-CD19 Microbeads (Miltenyi Biotec), according to the MiniMacs protocol (20 μl Microbeads + 80 μl PBS, for 107 cells). .. After the isolation procedure was ended, cells were maintained in PBS for 3 h at 4°C to minimize potential effects of anti-CD19 antibodies on B cell activation, then resuspended in TRIzol (Ambion-LifeTechnologies) for RNA extraction or in M-PER (Mammalian Protein Extraction Reagent, Thermo Scientific) for protein extraction.

Article Title: Inadequate Dietary Phosphorus Levels Cause Skeletal Anomalies and Alter Osteocalcin Gene Expression in Zebrafish
Article Snippet: RNA extraction was performed using the TRIzol method with 500 μL of TRIzol (GIBCO BRL, Waltham, MA, USA) for each sample to disrupt cells and release their contents. .. The samples were then incubated at 65 °C for 5 min and placed on ice for 1 min. To set the final solution, 0.50 mM of DTT (Dithiothreitol), 40 U of RNase, and 100 U of Super Script III were added.

Article Title: Oroxin B Induces Apoptosis by Down-Regulating MicroRNA-221 Resulting in the Inactivation of the PTEN/PI3K/AKT Pathway in Liver Cancer
Article Snippet: Microarray Analysis Total RNA was extracted from liver tissue, using TRIzol (Ambion, Carlsbad, CA, USA) according to the manufacturer’s instructions. .. After phosphatase treatment, 100 ng total RNA for each sample was incubated for 30 min at 37 °C.

Cell Culture:

Article Title: Stem cell transplantation impairs dendritic cell trafficking and herpesvirus immunity
Article Snippet: .. Total RNA was isolated from cultured cells or lung tissues by TRIzol (Invitrogen) according to manufacturer’s instructions. .. The relative amount of mRNA for viral genes or mouse genes was assayed by real-time reverse transcription PCR (real-time RT-PCR) using thermocycler ABI Prism 7000 (Applied Biosystems Inc.) using a previously described protocol ( ).

Expressing:

Article Title: Cell membrane coating for reducing nanoparticle-induced inflammatory responses to scaffold constructs
Article Snippet: Paragraph title: 2.7. Real-time PCR analysis of cytokine expression ... Fresh cells isolated from scaffolds were stored in TRIzol (Life Technologies), and RNA from cells was isolated using TRIzol reagent and TURBO DNA-free Kit (Invitrogen) according to manufacturer instructions.

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Article Snippet: Paragraph title: 4.6. Gene Expression ... RNA extraction was performed using the TRIzol method with 500 μL of TRIzol (GIBCO BRL, Waltham, MA, USA) for each sample to disrupt cells and release their contents.

Article Title: A vitamin C-derived DNA modification catalyzed by an algal TET homolog
Article Snippet: Total RNA was extracted from C. reinhardtii using Trizol™ (Thermo) according to the instruction manual. .. To measure the gene expression level, quantitative RT-PCR was performed using CFXP6™ Real-Time PCR with SYBR Premix™ Ex Taq (Tli RNaseH Plus, Takara).

Modification:

Article Title: Baicalein Suppresses Stem Cell-Like Characteristics in Radio- and Chemoresistant MDA-MB-231 Human Breast Cancer Cells through Up-Regulation of IFIT2
Article Snippet: .. Reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), insulin, bovine serum albumin (BSA), βFGF, EGF, B-27 supplement, 100 × trypsin/EDTA, 10 × streptomycin/antibiotics, and TRIzol were purchased from Gibco (Gaithersburg, MD, USA), except for TRIzol (Ambion, Austin, TX, USA). .. Baicalein, luteolin, myricetin, kaempferol, rutin, quercetin, HEPES, Adriamycin (doxorubicin), propidium iodide (PI), Hoechst 33342 dye, 2′,7′-dichlorofluorescin diacetate (H2 DCF-DA), and RNase A were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

Over Expression:

Article Title: HHIP overexpression inhibits the proliferation, migration and invasion of non-small cell lung cancer
Article Snippet: .. Experimental materials and reagents Human normal lung cell line (BEAS-2B) and NSCLC cell lines (H1975, H358, H226 and HCC827) were purchased from ATCC (USA); RPMI-1640 medium, fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco (USA); Trizol and LipofectamineTM 2000 were purchased from ThermoFisher (USA); pEGFP overexpression plasmid was purchased from Shanghai Beinuo Biotechnology Co., Ltd.; CCK-8 kit was purchased from Shanghai Beyotime Biotechnology Co., Ltd.; cDNA PrimeScript RT kit and SYBR-Green PCR Master Mix kit were purchased from Dalian TaKaRa Co., Ltd.; Transwell chamber was purchased from Milipore (USA); artificial Matrigel was purchased from BD Biosciences (USA). .. ABI 7500 fluorescence quantitative PCR was purchased from Applied Biosystems (USA).

Hybridization:

Article Title: Oroxin B Induces Apoptosis by Down-Regulating MicroRNA-221 Resulting in the Inactivation of the PTEN/PI3K/AKT Pathway in Liver Cancer
Article Snippet: Microarray Analysis Total RNA was extracted from liver tissue, using TRIzol (Ambion, Carlsbad, CA, USA) according to the manufacturer’s instructions. .. Samples were dried with vacuum concentrator, approximately 2 to 3 h at 55 °C, and then performed assemble hybridization.

Concentration Assay:

Article Title: Cell membrane coating for reducing nanoparticle-induced inflammatory responses to scaffold constructs
Article Snippet: Fresh cells isolated from scaffolds were stored in TRIzol (Life Technologies), and RNA from cells was isolated using TRIzol reagent and TURBO DNA-free Kit (Invitrogen) according to manufacturer instructions. .. The concentration of RNA was quantified using the NanoDrop 1000 Spectro-photomer (Thermo Scientific). cDNA was prepared according to the protocol provided by the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). qPCR was carried out using PowerUp SYBR Green Master Mix (Applied Biosystems) on QuantStudio 6 Flex Real-Time PCR System (Applied Biosystems).

Infection:

Article Title: Ifenprodil and Flavopiridol Identified by Genomewide RNA Interference Screening as Effective Drugs To Ameliorate Murine Acute Lung Injury after Influenza A H5N1 Virus Infection
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Polymerase Chain Reaction:

Article Title: HHIP overexpression inhibits the proliferation, migration and invasion of non-small cell lung cancer
Article Snippet: .. Experimental materials and reagents Human normal lung cell line (BEAS-2B) and NSCLC cell lines (H1975, H358, H226 and HCC827) were purchased from ATCC (USA); RPMI-1640 medium, fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco (USA); Trizol and LipofectamineTM 2000 were purchased from ThermoFisher (USA); pEGFP overexpression plasmid was purchased from Shanghai Beinuo Biotechnology Co., Ltd.; CCK-8 kit was purchased from Shanghai Beyotime Biotechnology Co., Ltd.; cDNA PrimeScript RT kit and SYBR-Green PCR Master Mix kit were purchased from Dalian TaKaRa Co., Ltd.; Transwell chamber was purchased from Milipore (USA); artificial Matrigel was purchased from BD Biosciences (USA). .. ABI 7500 fluorescence quantitative PCR was purchased from Applied Biosystems (USA).

Article Title: Stem cell transplantation impairs dendritic cell trafficking and herpesvirus immunity
Article Snippet: Total RNA was isolated from cultured cells or lung tissues by TRIzol (Invitrogen) according to manufacturer’s instructions. .. The relative amount of mRNA for viral genes or mouse genes was assayed by real-time reverse transcription PCR (real-time RT-PCR) using thermocycler ABI Prism 7000 (Applied Biosystems Inc.) using a previously described protocol ( ).

Radioactivity:

Article Title: In vivo BiFC analysis of Y14 and NXF1 mRNA export complexes: preferential localization within and around SC35 domains
Article Snippet: RNA purification with TRIzol After the immunoprecipitation, 1 ml TRIzol (Invitrogen) was added, mixed, and incubated at RT for 5 min. 200 μl chloroform was added, mixed, and incubated at RT for 15 min. .. Radioactivity was exposed on BioMax MS film (Kodak).

RNA Sequencing Assay:

Article Title: Ifenprodil and Flavopiridol Identified by Genomewide RNA Interference Screening as Effective Drugs To Ameliorate Murine Acute Lung Injury after Influenza A H5N1 Virus Infection
Article Snippet: Paragraph title: RNA sequencing and data analysis. ... Total RNA was isolated from lung tissue and blood using TRIzol (Invitrogen, CA, USA).

Fluorescence:

Article Title: HHIP overexpression inhibits the proliferation, migration and invasion of non-small cell lung cancer
Article Snippet: Experimental materials and reagents Human normal lung cell line (BEAS-2B) and NSCLC cell lines (H1975, H358, H226 and HCC827) were purchased from ATCC (USA); RPMI-1640 medium, fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco (USA); Trizol and LipofectamineTM 2000 were purchased from ThermoFisher (USA); pEGFP overexpression plasmid was purchased from Shanghai Beinuo Biotechnology Co., Ltd.; CCK-8 kit was purchased from Shanghai Beyotime Biotechnology Co., Ltd.; cDNA PrimeScript RT kit and SYBR-Green PCR Master Mix kit were purchased from Dalian TaKaRa Co., Ltd.; Transwell chamber was purchased from Milipore (USA); artificial Matrigel was purchased from BD Biosciences (USA). .. ABI 7500 fluorescence quantitative PCR was purchased from Applied Biosystems (USA).

Isolation:

Article Title: Obesity induces pro-inflammatory B cells and impairs B cell function in old mice
Article Snippet: .. After the isolation procedure was ended, cells were maintained in PBS for 3 h at 4°C to minimize potential effects of anti-CD19 antibodies on B cell activation, then resuspended in TRIzol (Ambion-LifeTechnologies) for RNA extraction or in M-PER (Mammalian Protein Extraction Reagent, Thermo Scientific) for protein extraction. .. Splenic B cells were cultured in complete medium (RPMI 1640, supplemented with 10% FCS, 10 μg/ml gentamicin, 2 × 10−5 M 2-ME, and 2 mM L-glutamine).

Article Title: Cell membrane coating for reducing nanoparticle-induced inflammatory responses to scaffold constructs
Article Snippet: .. Fresh cells isolated from scaffolds were stored in TRIzol (Life Technologies), and RNA from cells was isolated using TRIzol reagent and TURBO DNA-free Kit (Invitrogen) according to manufacturer instructions. .. The concentration of RNA was quantified using the NanoDrop 1000 Spectro-photomer (Thermo Scientific). cDNA was prepared according to the protocol provided by the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). qPCR was carried out using PowerUp SYBR Green Master Mix (Applied Biosystems) on QuantStudio 6 Flex Real-Time PCR System (Applied Biosystems).

Article Title: Ifenprodil and Flavopiridol Identified by Genomewide RNA Interference Screening as Effective Drugs To Ameliorate Murine Acute Lung Injury after Influenza A H5N1 Virus Infection
Article Snippet: .. Total RNA was isolated from lung tissue and blood using TRIzol (Invitrogen, CA, USA). .. High-throughput strand-specific RNA sequencing was performed using an Illumina HiSeq 2500 platform (Berry Genomics, Beijing, China).

Article Title: The emergent landscape of the mouse gut endoderm at single-cell resolution
Article Snippet: .. Total RNA was isolated from whole 13ss stage wild-type embryos using Trizol (Invitrogen). ..

Article Title: Stem cell transplantation impairs dendritic cell trafficking and herpesvirus immunity
Article Snippet: .. Total RNA was isolated from cultured cells or lung tissues by TRIzol (Invitrogen) according to manufacturer’s instructions. .. The relative amount of mRNA for viral genes or mouse genes was assayed by real-time reverse transcription PCR (real-time RT-PCR) using thermocycler ABI Prism 7000 (Applied Biosystems Inc.) using a previously described protocol ( ).

Labeling:

Article Title: Oroxin B Induces Apoptosis by Down-Regulating MicroRNA-221 Resulting in the Inactivation of the PTEN/PI3K/AKT Pathway in Liver Cancer
Article Snippet: Microarray Analysis Total RNA was extracted from liver tissue, using TRIzol (Ambion, Carlsbad, CA, USA) according to the manufacturer’s instructions. .. The mixture with DMSO was incubated at 16 °C for 2 h for assemble labeling reaction.

Purification:

Article Title: In vivo BiFC analysis of Y14 and NXF1 mRNA export complexes: preferential localization within and around SC35 domains
Article Snippet: .. RNA purification with TRIzol After the immunoprecipitation, 1 ml TRIzol (Invitrogen) was added, mixed, and incubated at RT for 5 min. 200 μl chloroform was added, mixed, and incubated at RT for 15 min. ..

Article Title: Obesity induces pro-inflammatory B cells and impairs B cell function in old mice
Article Snippet: At the end of the purification procedure, cells were 90–95% CD19-positive by cytofluorimetric analysis. .. After the isolation procedure was ended, cells were maintained in PBS for 3 h at 4°C to minimize potential effects of anti-CD19 antibodies on B cell activation, then resuspended in TRIzol (Ambion-LifeTechnologies) for RNA extraction or in M-PER (Mammalian Protein Extraction Reagent, Thermo Scientific) for protein extraction.

Article Title: Expression of T-plastin, FoxP3 and other tumor-associated markers by leukemic T-cells of cutaneous T-cell lymphoma
Article Snippet: .. For qPCR analysis, mRNA was extracted and purified with Trizol from lysate of 5 – 10×105 thawed and washed cells with 500 μL Trizol (Invitrogen, Carlsbad, CA) reagent. ..

Sequencing:

Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy
Article Snippet: RNA Extraction and Quantitative Real-Time PCR Total RNA was extracted using TRIzol (Thermofisher Scientific) and 1 μg total RNA was used to synthesize cDNA with SuperScript III Reverse Transcriptase (Thermofisher Scientific). .. Quantitative real-time (qRT)-PCR was performed in duplicate using SYBR Green Master Mix (Applied BioSystems) and an ABI PRIZM 7700 with Sequence Detector v1.7 (Applied Biosystems).

Protein Extraction:

Article Title: Obesity induces pro-inflammatory B cells and impairs B cell function in old mice
Article Snippet: .. After the isolation procedure was ended, cells were maintained in PBS for 3 h at 4°C to minimize potential effects of anti-CD19 antibodies on B cell activation, then resuspended in TRIzol (Ambion-LifeTechnologies) for RNA extraction or in M-PER (Mammalian Protein Extraction Reagent, Thermo Scientific) for protein extraction. .. Splenic B cells were cultured in complete medium (RPMI 1640, supplemented with 10% FCS, 10 μg/ml gentamicin, 2 × 10−5 M 2-ME, and 2 mM L-glutamine).

Quantitative RT-PCR:

Article Title: External quality assessment for PML‐RARα detection in acute promyelocytic leukemia: Findings and summary. External quality assessment for PML‐RARα detection in acute promyelocytic leukemia: Findings and summary
Article Snippet: Total RNA was extracted by TRIzol reagent and spin column, quantified using NanoDrop 2000c (Thermo Fisher). .. Using the one‐step or two‐step RT‐qPCR method, qualitative and quantitative detection of PML‐RARα FG and CG was performed by the manufacturer's instructions on ABI 7500 Instrument (Applied Biosystems).

Article Title: A vitamin C-derived DNA modification catalyzed by an algal TET homolog
Article Snippet: Total RNA was extracted from C. reinhardtii using Trizol™ (Thermo) according to the instruction manual. .. To measure the gene expression level, quantitative RT-PCR was performed using CFXP6™ Real-Time PCR with SYBR Premix™ Ex Taq (Tli RNaseH Plus, Takara).

Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy
Article Snippet: RNA Extraction and Quantitative Real-Time PCR Total RNA was extracted using TRIzol (Thermofisher Scientific) and 1 μg total RNA was used to synthesize cDNA with SuperScript III Reverse Transcriptase (Thermofisher Scientific). .. Quantitative real-time (qRT)-PCR was performed in duplicate using SYBR Green Master Mix (Applied BioSystems) and an ABI PRIZM 7700 with Sequence Detector v1.7 (Applied Biosystems).

Article Title: Stem cell transplantation impairs dendritic cell trafficking and herpesvirus immunity
Article Snippet: Total RNA was isolated from cultured cells or lung tissues by TRIzol (Invitrogen) according to manufacturer’s instructions. .. The relative amount of mRNA for viral genes or mouse genes was assayed by real-time reverse transcription PCR (real-time RT-PCR) using thermocycler ABI Prism 7000 (Applied Biosystems Inc.) using a previously described protocol ( ).

Mouse Assay:

Article Title: Obesity induces pro-inflammatory B cells and impairs B cell function in old mice
Article Snippet: B cells were isolated from the spleens or from the SVF of young and old mice after a 20 min incubation at 4°C with anti-CD19 Microbeads (Miltenyi Biotec), according to the MiniMacs protocol (20 μl Microbeads + 80 μl PBS, for 107 cells). .. After the isolation procedure was ended, cells were maintained in PBS for 3 h at 4°C to minimize potential effects of anti-CD19 antibodies on B cell activation, then resuspended in TRIzol (Ambion-LifeTechnologies) for RNA extraction or in M-PER (Mammalian Protein Extraction Reagent, Thermo Scientific) for protein extraction.

Article Title: Transcriptome analyses of adult mouse brain reveal enrichment of lncRNAs in specific brain regions and neuronal populations
Article Snippet: 8–12 week old male C57Bl/J6 mice (Jackson Laboratories) were sacrificed and the brain was removed from the skull and placed in ice-cold HEPES HBSS (HHBSS: 1× Hank's basal salt solution, 2.5 mm HEPES-KOH, pH 7.4, 35 mm glucose, and 4 mm NaHCO3). .. The hippocampus and PFC was dissected and homogenized in Trizol using a glass-Teflon homogenizer containing 1 ml of ice-cold Trizol (Life technologies, USA).

Article Title: Ifenprodil and Flavopiridol Identified by Genomewide RNA Interference Screening as Effective Drugs To Ameliorate Murine Acute Lung Injury after Influenza A H5N1 Virus Infection
Article Snippet: Lung tissue and blood from mice treated with drug or vehicle were obtained at 3 days after H5N1 infection. .. Total RNA was isolated from lung tissue and blood using TRIzol (Invitrogen, CA, USA).

Plasmid Preparation:

Article Title: HHIP overexpression inhibits the proliferation, migration and invasion of non-small cell lung cancer
Article Snippet: .. Experimental materials and reagents Human normal lung cell line (BEAS-2B) and NSCLC cell lines (H1975, H358, H226 and HCC827) were purchased from ATCC (USA); RPMI-1640 medium, fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco (USA); Trizol and LipofectamineTM 2000 were purchased from ThermoFisher (USA); pEGFP overexpression plasmid was purchased from Shanghai Beinuo Biotechnology Co., Ltd.; CCK-8 kit was purchased from Shanghai Beyotime Biotechnology Co., Ltd.; cDNA PrimeScript RT kit and SYBR-Green PCR Master Mix kit were purchased from Dalian TaKaRa Co., Ltd.; Transwell chamber was purchased from Milipore (USA); artificial Matrigel was purchased from BD Biosciences (USA). .. ABI 7500 fluorescence quantitative PCR was purchased from Applied Biosystems (USA).

Real-time Polymerase Chain Reaction:

Article Title: Cell membrane coating for reducing nanoparticle-induced inflammatory responses to scaffold constructs
Article Snippet: Paragraph title: 2.7. Real-time PCR analysis of cytokine expression ... Fresh cells isolated from scaffolds were stored in TRIzol (Life Technologies), and RNA from cells was isolated using TRIzol reagent and TURBO DNA-free Kit (Invitrogen) according to manufacturer instructions.

Article Title: A vitamin C-derived DNA modification catalyzed by an algal TET homolog
Article Snippet: Total RNA was extracted from C. reinhardtii using Trizol™ (Thermo) according to the instruction manual. .. To measure the gene expression level, quantitative RT-PCR was performed using CFXP6™ Real-Time PCR with SYBR Premix™ Ex Taq (Tli RNaseH Plus, Takara).

Article Title: Baicalein Suppresses Stem Cell-Like Characteristics in Radio- and Chemoresistant MDA-MB-231 Human Breast Cancer Cells through Up-Regulation of IFIT2
Article Snippet: Reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), insulin, bovine serum albumin (BSA), βFGF, EGF, B-27 supplement, 100 × trypsin/EDTA, 10 × streptomycin/antibiotics, and TRIzol were purchased from Gibco (Gaithersburg, MD, USA), except for TRIzol (Ambion, Austin, TX, USA). .. TOPreal qPCR preMIX was obtained from Enzynomics (Daejeon, Korea).

Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy
Article Snippet: .. RNA Extraction and Quantitative Real-Time PCR Total RNA was extracted using TRIzol (Thermofisher Scientific) and 1 μg total RNA was used to synthesize cDNA with SuperScript III Reverse Transcriptase (Thermofisher Scientific). .. Quantitative real-time (qRT)-PCR was performed in duplicate using SYBR Green Master Mix (Applied BioSystems) and an ABI PRIZM 7700 with Sequence Detector v1.7 (Applied Biosystems).

Article Title: Stem cell transplantation impairs dendritic cell trafficking and herpesvirus immunity
Article Snippet: Paragraph title: Quantitative PCR. ... Total RNA was isolated from cultured cells or lung tissues by TRIzol (Invitrogen) according to manufacturer’s instructions.

Article Title: Expression of T-plastin, FoxP3 and other tumor-associated markers by leukemic T-cells of cutaneous T-cell lymphoma
Article Snippet: .. For qPCR analysis, mRNA was extracted and purified with Trizol from lysate of 5 – 10×105 thawed and washed cells with 500 μL Trizol (Invitrogen, Carlsbad, CA) reagent. ..

RNA Extraction:

Article Title: Obesity induces pro-inflammatory B cells and impairs B cell function in old mice
Article Snippet: .. After the isolation procedure was ended, cells were maintained in PBS for 3 h at 4°C to minimize potential effects of anti-CD19 antibodies on B cell activation, then resuspended in TRIzol (Ambion-LifeTechnologies) for RNA extraction or in M-PER (Mammalian Protein Extraction Reagent, Thermo Scientific) for protein extraction. .. Splenic B cells were cultured in complete medium (RPMI 1640, supplemented with 10% FCS, 10 μg/ml gentamicin, 2 × 10−5 M 2-ME, and 2 mM L-glutamine).

Article Title: Inadequate Dietary Phosphorus Levels Cause Skeletal Anomalies and Alter Osteocalcin Gene Expression in Zebrafish
Article Snippet: .. RNA extraction was performed using the TRIzol method with 500 μL of TRIzol (GIBCO BRL, Waltham, MA, USA) for each sample to disrupt cells and release their contents. .. The extraction product was visualized on 1% agarose gel and quantified using a NanoDrop instrument (ND-1000 Spectrophotometer, UNESP Botucatu, Brazil).

Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy
Article Snippet: .. RNA Extraction and Quantitative Real-Time PCR Total RNA was extracted using TRIzol (Thermofisher Scientific) and 1 μg total RNA was used to synthesize cDNA with SuperScript III Reverse Transcriptase (Thermofisher Scientific). .. Quantitative real-time (qRT)-PCR was performed in duplicate using SYBR Green Master Mix (Applied BioSystems) and an ABI PRIZM 7700 with Sequence Detector v1.7 (Applied Biosystems).

Agarose Gel Electrophoresis:

Article Title: Inadequate Dietary Phosphorus Levels Cause Skeletal Anomalies and Alter Osteocalcin Gene Expression in Zebrafish
Article Snippet: RNA extraction was performed using the TRIzol method with 500 μL of TRIzol (GIBCO BRL, Waltham, MA, USA) for each sample to disrupt cells and release their contents. .. The extraction product was visualized on 1% agarose gel and quantified using a NanoDrop instrument (ND-1000 Spectrophotometer, UNESP Botucatu, Brazil).

Next-Generation Sequencing:

Article Title: Transcriptome analyses of adult mouse brain reveal enrichment of lncRNAs in specific brain regions and neuronal populations
Article Snippet: Paragraph title: RNA preparation for next generation sequencing (NGS) ... The hippocampus and PFC was dissected and homogenized in Trizol using a glass-Teflon homogenizer containing 1 ml of ice-cold Trizol (Life technologies, USA).

CCK-8 Assay:

Article Title: HHIP overexpression inhibits the proliferation, migration and invasion of non-small cell lung cancer
Article Snippet: .. Experimental materials and reagents Human normal lung cell line (BEAS-2B) and NSCLC cell lines (H1975, H358, H226 and HCC827) were purchased from ATCC (USA); RPMI-1640 medium, fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco (USA); Trizol and LipofectamineTM 2000 were purchased from ThermoFisher (USA); pEGFP overexpression plasmid was purchased from Shanghai Beinuo Biotechnology Co., Ltd.; CCK-8 kit was purchased from Shanghai Beyotime Biotechnology Co., Ltd.; cDNA PrimeScript RT kit and SYBR-Green PCR Master Mix kit were purchased from Dalian TaKaRa Co., Ltd.; Transwell chamber was purchased from Milipore (USA); artificial Matrigel was purchased from BD Biosciences (USA). .. ABI 7500 fluorescence quantitative PCR was purchased from Applied Biosystems (USA).

Spectrophotometry:

Article Title: Inadequate Dietary Phosphorus Levels Cause Skeletal Anomalies and Alter Osteocalcin Gene Expression in Zebrafish
Article Snippet: RNA extraction was performed using the TRIzol method with 500 μL of TRIzol (GIBCO BRL, Waltham, MA, USA) for each sample to disrupt cells and release their contents. .. The extraction product was visualized on 1% agarose gel and quantified using a NanoDrop instrument (ND-1000 Spectrophotometer, UNESP Botucatu, Brazil).

Activation Assay:

Article Title: Obesity induces pro-inflammatory B cells and impairs B cell function in old mice
Article Snippet: .. After the isolation procedure was ended, cells were maintained in PBS for 3 h at 4°C to minimize potential effects of anti-CD19 antibodies on B cell activation, then resuspended in TRIzol (Ambion-LifeTechnologies) for RNA extraction or in M-PER (Mammalian Protein Extraction Reagent, Thermo Scientific) for protein extraction. .. Splenic B cells were cultured in complete medium (RPMI 1640, supplemented with 10% FCS, 10 μg/ml gentamicin, 2 × 10−5 M 2-ME, and 2 mM L-glutamine).

Immunoprecipitation:

Article Title: In vivo BiFC analysis of Y14 and NXF1 mRNA export complexes: preferential localization within and around SC35 domains
Article Snippet: .. RNA purification with TRIzol After the immunoprecipitation, 1 ml TRIzol (Invitrogen) was added, mixed, and incubated at RT for 5 min. 200 μl chloroform was added, mixed, and incubated at RT for 15 min. ..

High Throughput Screening Assay:

Article Title: Ifenprodil and Flavopiridol Identified by Genomewide RNA Interference Screening as Effective Drugs To Ameliorate Murine Acute Lung Injury after Influenza A H5N1 Virus Infection
Article Snippet: Total RNA was isolated from lung tissue and blood using TRIzol (Invitrogen, CA, USA). .. High-throughput strand-specific RNA sequencing was performed using an Illumina HiSeq 2500 platform (Berry Genomics, Beijing, China).

Fluorescence In Situ Hybridization:

Article Title: Inadequate Dietary Phosphorus Levels Cause Skeletal Anomalies and Alter Osteocalcin Gene Expression in Zebrafish
Article Snippet: For RNA extraction, whole fish samples were individually macerated in liquid nitrogen with a ceramic crucible support and pistil previously autoclaved and decontaminated with RNase AWAY® (Thermo Fisher Scientific, Waltham, MA, USA). .. RNA extraction was performed using the TRIzol method with 500 μL of TRIzol (GIBCO BRL, Waltham, MA, USA) for each sample to disrupt cells and release their contents.

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    Thermo Fisher trizol reagent
    Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 722 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    This system includes both TRIzol LS Reagent and Invitrogen Phasemaker Tubes TRIzol LS Reagent is a complete ready to use reagent for the isolation of high quality total RNA from
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