tb green premix ex taq ii  (TaKaRa)

 
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    Name:
    TB Green Premix Ex Taq II
    Description:
    TB Green Premix Ex Taq II Tli RNaseH Plus is a TB Green Master Mix that effectively suppresses amplification of primer dimers and off target products and is therefore recommended for real time PCR qPCR when high specificity is critical Use this product when primer design has been optimized but when an increase in specificity of the qPCR assay is still needed TB Green Premix Ex Taq II Tli RNase H Plus includes TB Green a reagent designed for intercalator based qPCR
    Catalog Number:
    rr820b
    Price:
    None
    Size:
    400 Rxns
    Category:
    TB Green Premix Ex Taq II Tli RNase H Plus qPCR with TB Green detection Real time PCR kits Real time PCR
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    Structured Review

    TaKaRa tb green premix ex taq ii
    TB Green Premix Ex Taq II Tli RNaseH Plus is a TB Green Master Mix that effectively suppresses amplification of primer dimers and off target products and is therefore recommended for real time PCR qPCR when high specificity is critical Use this product when primer design has been optimized but when an increase in specificity of the qPCR assay is still needed TB Green Premix Ex Taq II Tli RNase H Plus includes TB Green a reagent designed for intercalator based qPCR
    https://www.bioz.com/result/tb green premix ex taq ii/product/TaKaRa
    Average 99 stars, based on 204 article reviews
    Price from $9.99 to $1999.99
    tb green premix ex taq ii - by Bioz Stars, 2020-11
    99/100 stars

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    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: The novel estrogenic receptor GPR30 alleviates ischemic injury by inhibiting TLR4-mediated microglial inflammation
    Article Snippet: .. Real-time PCR was performed in a 25-μl reaction according to the manufacturer’s manual for SYBR Premix Ex Taq (Tli RNaseH Plus) (#RR820A, TaKaRa, Japan). .. The TLR4 mRNA primer sequences [TaKaRa Biotechnology (Dalian) Co., Ltd.] were as follows: F: 5′-GGGCCTAAACCCAGTCTGTTTG-3′, R: 5′-CTTCTGCCCGGTAAGGTCCA-3′.

    Article Title: Trifluoperazine induces apoptosis through the upregulation of Bax/Bcl-2 and downregulated phosphorylation of AKT in mesangial cells and improves renal function in lupus nephritis mice
    Article Snippet: .. The qPCR analysis was performed using SYBR Premix Ex Taq (2X) (RR820A; Takara Bio, Inc.). .. A total of 2 μ l cDNA ( < 100 ng), 10 μ l buffer, 0.3 μ l forward primer, 0.3 μ l reverse primer and double-distilled water (ddH2 O) ≤20 μ l. The qPCR thermal cycling protocol was programmed in the CFX96™ Real-Time PCR Detection system (Bio-Rad Laboratories, Inc.) and consisted of an initial denaturation step at 95°C for 30 sec, followed by 40 cycles of denaturation for 5 sec at 95°C, and annealing and extension for 30 sec at 56°C.

    Article Title: Tripterygium wilfordii mitigates hyperglycemia-induced upregulated Wnt/β-catenin expression and kidney injury in diabetic rats
    Article Snippet: .. Following the determination of RNA concentration and purity by an ultraviolet spectrophotometer (NanoDrop™ 2000; Thermo Fisher Scientific Inc., Waltham, MA, USA), RNA samples (5 µg/sample) were reverse transcribed into cDNA using a PrimeScript™ RT reagent kit (cat. no. RR047A; Takara Biotechnology Co., Ltd., Dalian, China). qPCR was performed using the SYBR-Green kit (cat. no. RR820A; Takara Biotechnology Co., Ltd.) and specific primers (Sangon Biotech Co., Ltd., Shanghai, China) in an Applied Biosystems StepOne Real-Time PCR machine (Applied Biosystems; Thermo Fisher Scientific Inc.). .. The sequences of primers and product lengths are presented in . qPCR was performed using SYBR® Premix Ex Taq™ II in a 20 µl reaction volume.

    Article Title: Cytosolic Internalization of Anti-DNA Antibodies by Human Monocytes Induces Production of Pro-inflammatory Cytokines Independently of the Tripartite Motif-Containing 21 (TRIM21)-Mediated Pathway
    Article Snippet: .. First-strand cDNA was synthesized using a PrimeScript™ RT reagent Kit (Takara, Kusatsu, Japan; cat# RR037A), and quantitative reverse transcription-PCR (qRT-PCR) was carried out using the SYBR Premix Ex™ Taq II kit (Takara; cat# RR820A) and an ABI 7300 qPCR instrument (Applied Biosystems). .. The primers used for real-time RT-PCR were as follows: human IL-8, 5′-GAAGGACCTAGGACGGAAGG-3′ (forward) and 5′-GGGTGG AAAGGTTTGGAGTATG-3′ (reverse); human TNF-α, 5′-CTGCTGCACTTTGGAGTGAT-3′ (forward) and 5′-AGATGATCTGACTGCCTGGG-3′ (reverse); β-actin, 5′-CATGTACGTTGCTATCCAGGC-3′ (forward) and 5′-CTCCTTAATGTCACGCACGAT-3′ (reverse).

    Synthesized:

    Article Title: Cytosolic Internalization of Anti-DNA Antibodies by Human Monocytes Induces Production of Pro-inflammatory Cytokines Independently of the Tripartite Motif-Containing 21 (TRIM21)-Mediated Pathway
    Article Snippet: .. First-strand cDNA was synthesized using a PrimeScript™ RT reagent Kit (Takara, Kusatsu, Japan; cat# RR037A), and quantitative reverse transcription-PCR (qRT-PCR) was carried out using the SYBR Premix Ex™ Taq II kit (Takara; cat# RR820A) and an ABI 7300 qPCR instrument (Applied Biosystems). .. The primers used for real-time RT-PCR were as follows: human IL-8, 5′-GAAGGACCTAGGACGGAAGG-3′ (forward) and 5′-GGGTGG AAAGGTTTGGAGTATG-3′ (reverse); human TNF-α, 5′-CTGCTGCACTTTGGAGTGAT-3′ (forward) and 5′-AGATGATCTGACTGCCTGGG-3′ (reverse); β-actin, 5′-CATGTACGTTGCTATCCAGGC-3′ (forward) and 5′-CTCCTTAATGTCACGCACGAT-3′ (reverse).

    Quantitative RT-PCR:

    Article Title: Long noncoding RNA SNHG6 functions as a competing endogenous RNA by sponging miR-181a-5p to regulate E2F5 expression in colorectal cancer
    Article Snippet: .. Total RNA was reversely transcribed into cDNA using GoScript RT System (Promega, Madison, WI, USA) following the manufacturer’s protocol. qRT-PCR was performed using SYBR Premix EX Taq™ II kit (RR820A; TaKaRa, Kusatsu, China) following the manufacturer’s instructions using ABI 7500 (Applied Biosystems, Foster City, CA, USA). ..

    Article Title: Cytosolic Internalization of Anti-DNA Antibodies by Human Monocytes Induces Production of Pro-inflammatory Cytokines Independently of the Tripartite Motif-Containing 21 (TRIM21)-Mediated Pathway
    Article Snippet: .. First-strand cDNA was synthesized using a PrimeScript™ RT reagent Kit (Takara, Kusatsu, Japan; cat# RR037A), and quantitative reverse transcription-PCR (qRT-PCR) was carried out using the SYBR Premix Ex™ Taq II kit (Takara; cat# RR820A) and an ABI 7300 qPCR instrument (Applied Biosystems). .. The primers used for real-time RT-PCR were as follows: human IL-8, 5′-GAAGGACCTAGGACGGAAGG-3′ (forward) and 5′-GGGTGG AAAGGTTTGGAGTATG-3′ (reverse); human TNF-α, 5′-CTGCTGCACTTTGGAGTGAT-3′ (forward) and 5′-AGATGATCTGACTGCCTGGG-3′ (reverse); β-actin, 5′-CATGTACGTTGCTATCCAGGC-3′ (forward) and 5′-CTCCTTAATGTCACGCACGAT-3′ (reverse).

    SYBR Green Assay:

    Article Title: Tripterygium wilfordii mitigates hyperglycemia-induced upregulated Wnt/β-catenin expression and kidney injury in diabetic rats
    Article Snippet: .. Following the determination of RNA concentration and purity by an ultraviolet spectrophotometer (NanoDrop™ 2000; Thermo Fisher Scientific Inc., Waltham, MA, USA), RNA samples (5 µg/sample) were reverse transcribed into cDNA using a PrimeScript™ RT reagent kit (cat. no. RR047A; Takara Biotechnology Co., Ltd., Dalian, China). qPCR was performed using the SYBR-Green kit (cat. no. RR820A; Takara Biotechnology Co., Ltd.) and specific primers (Sangon Biotech Co., Ltd., Shanghai, China) in an Applied Biosystems StepOne Real-Time PCR machine (Applied Biosystems; Thermo Fisher Scientific Inc.). .. The sequences of primers and product lengths are presented in . qPCR was performed using SYBR® Premix Ex Taq™ II in a 20 µl reaction volume.

    Concentration Assay:

    Article Title: Tripterygium wilfordii mitigates hyperglycemia-induced upregulated Wnt/β-catenin expression and kidney injury in diabetic rats
    Article Snippet: .. Following the determination of RNA concentration and purity by an ultraviolet spectrophotometer (NanoDrop™ 2000; Thermo Fisher Scientific Inc., Waltham, MA, USA), RNA samples (5 µg/sample) were reverse transcribed into cDNA using a PrimeScript™ RT reagent kit (cat. no. RR047A; Takara Biotechnology Co., Ltd., Dalian, China). qPCR was performed using the SYBR-Green kit (cat. no. RR820A; Takara Biotechnology Co., Ltd.) and specific primers (Sangon Biotech Co., Ltd., Shanghai, China) in an Applied Biosystems StepOne Real-Time PCR machine (Applied Biosystems; Thermo Fisher Scientific Inc.). .. The sequences of primers and product lengths are presented in . qPCR was performed using SYBR® Premix Ex Taq™ II in a 20 µl reaction volume.

    Spectrophotometry:

    Article Title: Tripterygium wilfordii mitigates hyperglycemia-induced upregulated Wnt/β-catenin expression and kidney injury in diabetic rats
    Article Snippet: .. Following the determination of RNA concentration and purity by an ultraviolet spectrophotometer (NanoDrop™ 2000; Thermo Fisher Scientific Inc., Waltham, MA, USA), RNA samples (5 µg/sample) were reverse transcribed into cDNA using a PrimeScript™ RT reagent kit (cat. no. RR047A; Takara Biotechnology Co., Ltd., Dalian, China). qPCR was performed using the SYBR-Green kit (cat. no. RR820A; Takara Biotechnology Co., Ltd.) and specific primers (Sangon Biotech Co., Ltd., Shanghai, China) in an Applied Biosystems StepOne Real-Time PCR machine (Applied Biosystems; Thermo Fisher Scientific Inc.). .. The sequences of primers and product lengths are presented in . qPCR was performed using SYBR® Premix Ex Taq™ II in a 20 µl reaction volume.

    Expressing:

    Article Title: Integration of GWAS and brain eQTL identifies FLOT1 as a risk gene for major depressive disorder
    Article Snippet: .. TB Green™ Premix Ex Taq™ II (Tli RNaseH Plus) (Takara, RR820A) was used to quantify the FLOT1 expression level. .. The QuantStudio™12 K Flex (Applied Biosystems) instrument was used to conduct the real-time quantitative PCR (qPCR).

    Polymerase Chain Reaction:

    Article Title: Comparison of the Oocyte Quality Derived from Two-Dimensional Follicle Culture Methods and Developmental Competence of In Vitro Grown and Matured Oocytes
    Article Snippet: .. Reactions were performed according to protocols included with the SYBR premix dimer eraser PCR kit (Takara Bio, RR820L). .. The PCR protocol used a denaturation step of 95°C for 10 min, followed by an amplification and quantification program that was repeated 40 times (95°C for 15 s and 60°C for 1 min) and a melting curve program (60–95°C with a heating rate of 0.34/s and continuous fluorescence measurement).

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    TaKaRa circrna 0079201
    <t>hsa_circRNA_0079201</t> significantly increases SMAD2 expression by suppressing miR-140-3p. SMAD2 (A) mRNA and (B) protein expression level was increased compared with that in the control group following overcircRNA_0079201. The expression of IHH, a SMAD2 target gene, was decreased compared with that in the control group following overcircRNA_0079201. (C and D) Though SMAD2 showed upregulation in the overcircRNA_0079201 group compared with the normal control group, no significant difference was observed in the SMAD2 mRNA or protein expression levels between the overcircRNA_ 0079201+ miR-140-3p mimic group and normal control group. Like SMAD2, IHH expression did not also demonstrate significant difference between the overcircRNA_ 0079201+ miR-140-3p mimic group and normal control group. This indicated hsa_circRNA_0079201 significantly increases SMAD2 expression and decreased IHH expression by suppressing miR-140-3p. The data are presented as the mean ± SD. n=3. *** P
    Circrna 0079201, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/circrna 0079201/product/TaKaRa
    Average 90 stars, based on 1 article reviews
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    circrna 0079201 - by Bioz Stars, 2020-11
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    99
    TaKaRa tb green premix ex taq ii mastermix
    Specificity of qPCR-ML (A) and qPCR-ama (B). The qPCR amplicons were run on a 2% agarose gel. Both qPCR assays were performed using three different PCR master mix: SYBR green PCR master mix, Diatheva (I); RT2 SYBR Green ROX FAST <t>Mastermix,</t> Qiagen (II); TB Green premix ex <t>Taq</t> II Mastermix, Takara Bio (III). For each master mix, DNA from L. ( L. ) infantum (0,006 ng/μl) , L. ( L. ) amazonensis (0,15 ng/μl) , Trypanosoma cruzi (0,1 ng/μl) and human DNA (30 ng/μl) were tested with the condition described in the manuscript. As negative control, a no template control was used for each qPCR run. M: ΦX174 DNA/ Bsu RI ( Hae III) Marker, 9 (ThermoFisher Scientific); NTC: no template control.
    Tb Green Premix Ex Taq Ii Mastermix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tb green premix ex taq ii mastermix/product/TaKaRa
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tb green premix ex taq ii mastermix - by Bioz Stars, 2020-11
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    99
    TaKaRa real time quantitative reverse transcription pcr qrt pcr
    Broad-spectrum antiviral activities of DHODH inhibitors. (A) Anti-Ebola replication efficacy. BSR-T7/5 cells were transfected with the EBOV mini-genome replication system (NP, VP35, VP30, MG, and L) in the presence of increasing concentrations of Teriflunomide, Brequinar, S312 and S416 respectively. Inhibitory effects of these compounds (EC 50 ) to EBOV mini-genome replication were determined using Bright-Glo Luciferase Assay (left-hand scale, red curve). CC 50 of compounds were determined by analyzing BSR-T7/5 cell viability using CellTiterGlo Assay (right-hand scale, green curve). The results are presented as a mean of at least two replicates ± SD. (B) Anti-Zika virus efficacy. Huh7 cells were infected with Zika virus (MOI=0.05) for 4 hours and then treated with increasing concentrations of compounds Teriflunomide, Brequinar, S312 and S416 respectively. The viral yields in cell supernatants were then quantified by <t>qRT-PCR</t> to reflect the replication efficiency of Zika virus. (C) Anti-SARS-CoV-2 virus efficacy. Aliquots of Vero E6 cells were seeded in 96-well plates and then infected with Beta CoV/Wuhan/WIV04/2019 at MOI of 0.03. At the same time, different concentrations of the compounds were added for co-culture. Cell supernatants were harvested 48 h.p.i. and RNA was extracted and quantified by qRT-PCR to determine the numbers of viral RNA copies. (D) Immuno-fluorescence assay of SARS-CoV-2-infected cells. Vero E6 cells were infected with SARS-CoV-2 under the same procedure of C. Cells were fixed and permeabilized for staining with anti-viral NP antibody, followed by staining with Alexa 488-labeled secondary antibody. Green represents infected cells. Nuclei were stained by DAPI, and the merge of NP and nuclei were shown. Scale bar, 400uM. The results (B, C) are presented as a mean of at least three replicates ± SD. Statistical analysis, One-way ANOVA for (B). NS, p > 0.05; *, p
    Real Time Quantitative Reverse Transcription Pcr Qrt Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time quantitative reverse transcription pcr qrt pcr/product/TaKaRa
    Average 99 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    real time quantitative reverse transcription pcr qrt pcr - by Bioz Stars, 2020-11
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    Image Search Results


    hsa_circRNA_0079201 significantly increases SMAD2 expression by suppressing miR-140-3p. SMAD2 (A) mRNA and (B) protein expression level was increased compared with that in the control group following overcircRNA_0079201. The expression of IHH, a SMAD2 target gene, was decreased compared with that in the control group following overcircRNA_0079201. (C and D) Though SMAD2 showed upregulation in the overcircRNA_0079201 group compared with the normal control group, no significant difference was observed in the SMAD2 mRNA or protein expression levels between the overcircRNA_ 0079201+ miR-140-3p mimic group and normal control group. Like SMAD2, IHH expression did not also demonstrate significant difference between the overcircRNA_ 0079201+ miR-140-3p mimic group and normal control group. This indicated hsa_circRNA_0079201 significantly increases SMAD2 expression and decreased IHH expression by suppressing miR-140-3p. The data are presented as the mean ± SD. n=3. *** P

    Journal: International Journal of Molecular Medicine

    Article Title: Hsa_circularRNA_0079201 suppresses chondrocyte proliferation and endochondral ossification by regulating the microRNA-140-3p/SMAD2 signaling pathway in idiopathic short stature

    doi: 10.3892/ijmm.2020.4737

    Figure Lengend Snippet: hsa_circRNA_0079201 significantly increases SMAD2 expression by suppressing miR-140-3p. SMAD2 (A) mRNA and (B) protein expression level was increased compared with that in the control group following overcircRNA_0079201. The expression of IHH, a SMAD2 target gene, was decreased compared with that in the control group following overcircRNA_0079201. (C and D) Though SMAD2 showed upregulation in the overcircRNA_0079201 group compared with the normal control group, no significant difference was observed in the SMAD2 mRNA or protein expression levels between the overcircRNA_ 0079201+ miR-140-3p mimic group and normal control group. Like SMAD2, IHH expression did not also demonstrate significant difference between the overcircRNA_ 0079201+ miR-140-3p mimic group and normal control group. This indicated hsa_circRNA_0079201 significantly increases SMAD2 expression and decreased IHH expression by suppressing miR-140-3p. The data are presented as the mean ± SD. n=3. *** P

    Article Snippet: The RT-qPCR protocol used for circRNA-0079201 (TB Green® Premix Ex Taq™ II; Takara Bio, Inc.) was as follows: Initial denaturation at 95°C for 10 min, followed by 40 cycles of 95°C for 10 sec, and 60°C for 34 sec. GAPDH was used as the internal control.

    Techniques: Expressing

    miR-140-3p is a hsa_circRNA_0079201 target gene. (A) miR-140-3p expression was significantly reduced after hsa_circRNA_0079201 was over-expressed in human chondrocytes. (B) The potential target binding sites between miR-140-3p and cirRNA-0079201. (C) Luciferase activity was repressed in human chondrocyte co-transfected with WT hsa_circRNA_0079201 and miR-140-3p mimics, while it was restored in cells co-transfected with Mut hsa_circRNA_0079201 and miR-140-3p mimics. (D) The co-localization of circRNA_0079201 and miR-140-3p in the human chondrocyte and neonatal femur growth plate of C57 mice was further validated using in situ hybridization. The data are presented as the mean ± SD. n=3. ** P

    Journal: International Journal of Molecular Medicine

    Article Title: Hsa_circularRNA_0079201 suppresses chondrocyte proliferation and endochondral ossification by regulating the microRNA-140-3p/SMAD2 signaling pathway in idiopathic short stature

    doi: 10.3892/ijmm.2020.4737

    Figure Lengend Snippet: miR-140-3p is a hsa_circRNA_0079201 target gene. (A) miR-140-3p expression was significantly reduced after hsa_circRNA_0079201 was over-expressed in human chondrocytes. (B) The potential target binding sites between miR-140-3p and cirRNA-0079201. (C) Luciferase activity was repressed in human chondrocyte co-transfected with WT hsa_circRNA_0079201 and miR-140-3p mimics, while it was restored in cells co-transfected with Mut hsa_circRNA_0079201 and miR-140-3p mimics. (D) The co-localization of circRNA_0079201 and miR-140-3p in the human chondrocyte and neonatal femur growth plate of C57 mice was further validated using in situ hybridization. The data are presented as the mean ± SD. n=3. ** P

    Article Snippet: The RT-qPCR protocol used for circRNA-0079201 (TB Green® Premix Ex Taq™ II; Takara Bio, Inc.) was as follows: Initial denaturation at 95°C for 10 min, followed by 40 cycles of 95°C for 10 sec, and 60°C for 34 sec. GAPDH was used as the internal control.

    Techniques: Expressing, Binding Assay, Luciferase, Activity Assay, Transfection, Mouse Assay, In Situ Hybridization

    OvercircRNA_0079201 expression significantly decreases human chondrocyte proliferation and causes cell cycle arrest. (A) OvercircRNA_0079201 was confirmed in patients with ISS and normal controls using RT-qPCR. (B) hsa_circRNA_0079201 mRNA expression level was not significantly different between samples treated with RNase R from the patients with ISS and the normal controls. (C) Linear IQCE was significantly decreased following RNase R treatment. (D) hsa_circRNA_0079201 mRNA expression level was significantly increased following transfection with overcircRNA_ 0079201. (E) The CCK-8 and (F) EdU assays showed that overcircRNA_0079201 significantly suppressed human chondrocyte proliferation. (G) Flow cytometry analysis revealed that the cells transfected with overcircRNA_ 0079201 were arrested in the G0/G1 phase. The data are presented as the mean ± SD. n=3. ** P

    Journal: International Journal of Molecular Medicine

    Article Title: Hsa_circularRNA_0079201 suppresses chondrocyte proliferation and endochondral ossification by regulating the microRNA-140-3p/SMAD2 signaling pathway in idiopathic short stature

    doi: 10.3892/ijmm.2020.4737

    Figure Lengend Snippet: OvercircRNA_0079201 expression significantly decreases human chondrocyte proliferation and causes cell cycle arrest. (A) OvercircRNA_0079201 was confirmed in patients with ISS and normal controls using RT-qPCR. (B) hsa_circRNA_0079201 mRNA expression level was not significantly different between samples treated with RNase R from the patients with ISS and the normal controls. (C) Linear IQCE was significantly decreased following RNase R treatment. (D) hsa_circRNA_0079201 mRNA expression level was significantly increased following transfection with overcircRNA_ 0079201. (E) The CCK-8 and (F) EdU assays showed that overcircRNA_0079201 significantly suppressed human chondrocyte proliferation. (G) Flow cytometry analysis revealed that the cells transfected with overcircRNA_ 0079201 were arrested in the G0/G1 phase. The data are presented as the mean ± SD. n=3. ** P

    Article Snippet: The RT-qPCR protocol used for circRNA-0079201 (TB Green® Premix Ex Taq™ II; Takara Bio, Inc.) was as follows: Initial denaturation at 95°C for 10 min, followed by 40 cycles of 95°C for 10 sec, and 60°C for 34 sec. GAPDH was used as the internal control.

    Techniques: Expressing, Quantitative RT-PCR, Transfection, CCK-8 Assay, Flow Cytometry

    OvercircRNA-0079201 inhibits chondrocyte hypertrophy and endochondral ossification. (A) The protein and mRNA expression level of RUNX2 and COL10A1 was decreased in cells transfected with over-circRNA-0079201, detected using western blot analysis and RT-qPCR, respectively. (B) Von Kossa staining showed the reduced mineralization in cells transfected with overcircRNA-0079201. The positive spots are indictaed using a red arrow. (C) ALP staining revealed that ALP activity was reduced in cells transfected with overcircRNA-0079201. The positive spots are indicated using a red arrow. The data are presented as the mean ± SD. n=3. *** P

    Journal: International Journal of Molecular Medicine

    Article Title: Hsa_circularRNA_0079201 suppresses chondrocyte proliferation and endochondral ossification by regulating the microRNA-140-3p/SMAD2 signaling pathway in idiopathic short stature

    doi: 10.3892/ijmm.2020.4737

    Figure Lengend Snippet: OvercircRNA-0079201 inhibits chondrocyte hypertrophy and endochondral ossification. (A) The protein and mRNA expression level of RUNX2 and COL10A1 was decreased in cells transfected with over-circRNA-0079201, detected using western blot analysis and RT-qPCR, respectively. (B) Von Kossa staining showed the reduced mineralization in cells transfected with overcircRNA-0079201. The positive spots are indictaed using a red arrow. (C) ALP staining revealed that ALP activity was reduced in cells transfected with overcircRNA-0079201. The positive spots are indicated using a red arrow. The data are presented as the mean ± SD. n=3. *** P

    Article Snippet: The RT-qPCR protocol used for circRNA-0079201 (TB Green® Premix Ex Taq™ II; Takara Bio, Inc.) was as follows: Initial denaturation at 95°C for 10 min, followed by 40 cycles of 95°C for 10 sec, and 60°C for 34 sec. GAPDH was used as the internal control.

    Techniques: Expressing, Transfection, Western Blot, Quantitative RT-PCR, Staining, Activity Assay

    Specificity of qPCR-ML (A) and qPCR-ama (B). The qPCR amplicons were run on a 2% agarose gel. Both qPCR assays were performed using three different PCR master mix: SYBR green PCR master mix, Diatheva (I); RT2 SYBR Green ROX FAST Mastermix, Qiagen (II); TB Green premix ex Taq II Mastermix, Takara Bio (III). For each master mix, DNA from L. ( L. ) infantum (0,006 ng/μl) , L. ( L. ) amazonensis (0,15 ng/μl) , Trypanosoma cruzi (0,1 ng/μl) and human DNA (30 ng/μl) were tested with the condition described in the manuscript. As negative control, a no template control was used for each qPCR run. M: ΦX174 DNA/ Bsu RI ( Hae III) Marker, 9 (ThermoFisher Scientific); NTC: no template control.

    Journal: Data in Brief

    Article Title: Data on the differentiation among Leishmania (Viannia) spp., Leishmania (Leishmania) infantum and Leishmania (Leishmania) amazonensis in Brazilian clinical samples using real-time PCR

    doi: 10.1016/j.dib.2019.104914

    Figure Lengend Snippet: Specificity of qPCR-ML (A) and qPCR-ama (B). The qPCR amplicons were run on a 2% agarose gel. Both qPCR assays were performed using three different PCR master mix: SYBR green PCR master mix, Diatheva (I); RT2 SYBR Green ROX FAST Mastermix, Qiagen (II); TB Green premix ex Taq II Mastermix, Takara Bio (III). For each master mix, DNA from L. ( L. ) infantum (0,006 ng/μl) , L. ( L. ) amazonensis (0,15 ng/μl) , Trypanosoma cruzi (0,1 ng/μl) and human DNA (30 ng/μl) were tested with the condition described in the manuscript. As negative control, a no template control was used for each qPCR run. M: ΦX174 DNA/ Bsu RI ( Hae III) Marker, 9 (ThermoFisher Scientific); NTC: no template control.

    Article Snippet: Three different PCR master mix were tested: SYBR green PCR master mix (Diatheva srl, Fano, Italy), RT2 SYBR Green ROX FAST Mastermix (Qiagen, Hilden, Germany), TB Green premix ex Taq II Mastermix (Takara Bio Europe, France).

    Techniques: Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Polymerase Chain Reaction, SYBR Green Assay, Negative Control, Marker

    Broad-spectrum antiviral activities of DHODH inhibitors. (A) Anti-Ebola replication efficacy. BSR-T7/5 cells were transfected with the EBOV mini-genome replication system (NP, VP35, VP30, MG, and L) in the presence of increasing concentrations of Teriflunomide, Brequinar, S312 and S416 respectively. Inhibitory effects of these compounds (EC 50 ) to EBOV mini-genome replication were determined using Bright-Glo Luciferase Assay (left-hand scale, red curve). CC 50 of compounds were determined by analyzing BSR-T7/5 cell viability using CellTiterGlo Assay (right-hand scale, green curve). The results are presented as a mean of at least two replicates ± SD. (B) Anti-Zika virus efficacy. Huh7 cells were infected with Zika virus (MOI=0.05) for 4 hours and then treated with increasing concentrations of compounds Teriflunomide, Brequinar, S312 and S416 respectively. The viral yields in cell supernatants were then quantified by qRT-PCR to reflect the replication efficiency of Zika virus. (C) Anti-SARS-CoV-2 virus efficacy. Aliquots of Vero E6 cells were seeded in 96-well plates and then infected with Beta CoV/Wuhan/WIV04/2019 at MOI of 0.03. At the same time, different concentrations of the compounds were added for co-culture. Cell supernatants were harvested 48 h.p.i. and RNA was extracted and quantified by qRT-PCR to determine the numbers of viral RNA copies. (D) Immuno-fluorescence assay of SARS-CoV-2-infected cells. Vero E6 cells were infected with SARS-CoV-2 under the same procedure of C. Cells were fixed and permeabilized for staining with anti-viral NP antibody, followed by staining with Alexa 488-labeled secondary antibody. Green represents infected cells. Nuclei were stained by DAPI, and the merge of NP and nuclei were shown. Scale bar, 400uM. The results (B, C) are presented as a mean of at least three replicates ± SD. Statistical analysis, One-way ANOVA for (B). NS, p > 0.05; *, p

    Journal: bioRxiv

    Article Title: Novel and potent inhibitors targeting DHODH, a rate-limiting enzyme in de novo pyrimidine biosynthesis, are broad-spectrum antiviral against RNA viruses including newly emerged coronavirus SARS-CoV-2

    doi: 10.1101/2020.03.11.983056

    Figure Lengend Snippet: Broad-spectrum antiviral activities of DHODH inhibitors. (A) Anti-Ebola replication efficacy. BSR-T7/5 cells were transfected with the EBOV mini-genome replication system (NP, VP35, VP30, MG, and L) in the presence of increasing concentrations of Teriflunomide, Brequinar, S312 and S416 respectively. Inhibitory effects of these compounds (EC 50 ) to EBOV mini-genome replication were determined using Bright-Glo Luciferase Assay (left-hand scale, red curve). CC 50 of compounds were determined by analyzing BSR-T7/5 cell viability using CellTiterGlo Assay (right-hand scale, green curve). The results are presented as a mean of at least two replicates ± SD. (B) Anti-Zika virus efficacy. Huh7 cells were infected with Zika virus (MOI=0.05) for 4 hours and then treated with increasing concentrations of compounds Teriflunomide, Brequinar, S312 and S416 respectively. The viral yields in cell supernatants were then quantified by qRT-PCR to reflect the replication efficiency of Zika virus. (C) Anti-SARS-CoV-2 virus efficacy. Aliquots of Vero E6 cells were seeded in 96-well plates and then infected with Beta CoV/Wuhan/WIV04/2019 at MOI of 0.03. At the same time, different concentrations of the compounds were added for co-culture. Cell supernatants were harvested 48 h.p.i. and RNA was extracted and quantified by qRT-PCR to determine the numbers of viral RNA copies. (D) Immuno-fluorescence assay of SARS-CoV-2-infected cells. Vero E6 cells were infected with SARS-CoV-2 under the same procedure of C. Cells were fixed and permeabilized for staining with anti-viral NP antibody, followed by staining with Alexa 488-labeled secondary antibody. Green represents infected cells. Nuclei were stained by DAPI, and the merge of NP and nuclei were shown. Scale bar, 400uM. The results (B, C) are presented as a mean of at least three replicates ± SD. Statistical analysis, One-way ANOVA for (B). NS, p > 0.05; *, p

    Article Snippet: The number of viral copies was detected by real-time quantitative reverse transcription PCR (qRT-PCR) (Takara Cat no.RR820A) to reflect the replication efficiency of the virus.

    Techniques: Transfection, Luciferase, Infection, Quantitative RT-PCR, Co-Culture Assay, Fluorescence, Staining, Labeling

    Significant upregulation of FLOT1 in peripheral blood of first-episode drug-naive MDD cases compared with controls. Quantitative PCR showed that FLOT1 was significantly upregulated ( P = 1.01 × 10 −7 ) in peripheral blood of first-episode drug-naive MDD cases compared with controls. Expression of FLOT1 was quantified in 46 controls and 50 MDD cases. Two-tailed Student’s t -test was used to compare if the different was significant ( P

    Journal: Neuropsychopharmacology

    Article Title: Integration of GWAS and brain eQTL identifies FLOT1 as a risk gene for major depressive disorder

    doi: 10.1038/s41386-019-0345-4

    Figure Lengend Snippet: Significant upregulation of FLOT1 in peripheral blood of first-episode drug-naive MDD cases compared with controls. Quantitative PCR showed that FLOT1 was significantly upregulated ( P = 1.01 × 10 −7 ) in peripheral blood of first-episode drug-naive MDD cases compared with controls. Expression of FLOT1 was quantified in 46 controls and 50 MDD cases. Two-tailed Student’s t -test was used to compare if the different was significant ( P

    Article Snippet: TB Green™ Premix Ex Taq™ II (Tli RNaseH Plus) (Takara, RR820A) was used to quantify the FLOT1 expression level.

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Two Tailed Test

    FLOT1 is widely expressed in diverse human tissues. RNA sequencing-based expression data from GTEx was used to explore FLOT1 expression. FLOT1 was abundantly expressed in different human tissues, with the highest expression level in blood

    Journal: Neuropsychopharmacology

    Article Title: Integration of GWAS and brain eQTL identifies FLOT1 as a risk gene for major depressive disorder

    doi: 10.1038/s41386-019-0345-4

    Figure Lengend Snippet: FLOT1 is widely expressed in diverse human tissues. RNA sequencing-based expression data from GTEx was used to explore FLOT1 expression. FLOT1 was abundantly expressed in different human tissues, with the highest expression level in blood

    Article Snippet: TB Green™ Premix Ex Taq™ II (Tli RNaseH Plus) (Takara, RR820A) was used to quantify the FLOT1 expression level.

    Techniques: RNA Sequencing Assay, Expressing

    Expression of FLOT1 in different cell types of human and mouse brain. a FLOT1 was expressed in different cell types (including neuron, microglia, oligodendrocytes, and astrocytes) of human brain. b FLOT1 was abundantly expressed in different cell types of mouse brain, with the relatively high expression level in neurons and microglia cells. c , d Expression of FLOT1 was relatively low at early developmental stage. With the progress of development , FLOT1 expression was gradually increased, and peaked at adult stage. c Expression data from the BrainSpan was used for plotting. d Expression data from the BrainCloud was used for plotting

    Journal: Neuropsychopharmacology

    Article Title: Integration of GWAS and brain eQTL identifies FLOT1 as a risk gene for major depressive disorder

    doi: 10.1038/s41386-019-0345-4

    Figure Lengend Snippet: Expression of FLOT1 in different cell types of human and mouse brain. a FLOT1 was expressed in different cell types (including neuron, microglia, oligodendrocytes, and astrocytes) of human brain. b FLOT1 was abundantly expressed in different cell types of mouse brain, with the relatively high expression level in neurons and microglia cells. c , d Expression of FLOT1 was relatively low at early developmental stage. With the progress of development , FLOT1 expression was gradually increased, and peaked at adult stage. c Expression data from the BrainSpan was used for plotting. d Expression data from the BrainCloud was used for plotting

    Article Snippet: TB Green™ Premix Ex Taq™ II (Tli RNaseH Plus) (Takara, RR820A) was used to quantify the FLOT1 expression level.

    Techniques: Expressing

    Significant upregulation of FLOT1 in brains of MDD cases compared with controls. a , c Compared with controls, FLOT1 was significantly upregulated in the dorsolateral prefrontal cortex (DLPFC) and anterior insula (AINS) of MDD cases. b , d HLA-C was significantly upregulated in the dorsolateral prefrontal cortex (DLPFC) and anterior insula (AINS) of MDD cases. One-tailed Student’s t -test was used to compare if the different was significant ( P

    Journal: Neuropsychopharmacology

    Article Title: Integration of GWAS and brain eQTL identifies FLOT1 as a risk gene for major depressive disorder

    doi: 10.1038/s41386-019-0345-4

    Figure Lengend Snippet: Significant upregulation of FLOT1 in brains of MDD cases compared with controls. a , c Compared with controls, FLOT1 was significantly upregulated in the dorsolateral prefrontal cortex (DLPFC) and anterior insula (AINS) of MDD cases. b , d HLA-C was significantly upregulated in the dorsolateral prefrontal cortex (DLPFC) and anterior insula (AINS) of MDD cases. One-tailed Student’s t -test was used to compare if the different was significant ( P

    Article Snippet: TB Green™ Premix Ex Taq™ II (Tli RNaseH Plus) (Takara, RR820A) was used to quantify the FLOT1 expression level.

    Techniques: One-tailed Test