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Bio-Rad qrt pcr
Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 95/100, based on 5746 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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qrt pcr - by Bioz Stars, 2021-01
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Real-time Polymerase Chain Reaction:

Article Title: RNA-seq analysis reveals the role of red light in resistance against Pseudomonas syringae pv. tomato DC3000 in tomato plants
Article Snippet: .. Verification of RNA-seq results by real-time PCR (qRT-PCR) To synthesis cDNA, total RNA from diluted stocks of the same RNA that was subjected to RNA-seq was used in each reverse transcription reaction using the ReverTra Ace qRT-PCR Kit (Toyobo, Japan). qRT-PCR was performed using SYBR-Green chemistry and the iCycler iQ™ Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) [ ]. ..

SYBR Green Assay:

Article Title: RNA-seq analysis reveals the role of red light in resistance against Pseudomonas syringae pv. tomato DC3000 in tomato plants
Article Snippet: .. Verification of RNA-seq results by real-time PCR (qRT-PCR) To synthesis cDNA, total RNA from diluted stocks of the same RNA that was subjected to RNA-seq was used in each reverse transcription reaction using the ReverTra Ace qRT-PCR Kit (Toyobo, Japan). qRT-PCR was performed using SYBR-Green chemistry and the iCycler iQ™ Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) [ ]. ..

other:

Article Title: Attenuation of Tick-Borne Encephalitis Virus Using Large-Scale Random Codon Re-encoding
Article Snippet: The mix content for a final volume of 25μL per sample, was as follows: a standard quantity of 2x of PCR Mastermix and Enzymes, both primers (final concentration: 0.4μM), probe (final concentration: 0.1μM) and 4μL of extracted nucleic acids. qRT-PCR were performed on CFX96 Real-Time System/C1000 Touch Thermal Cycler (Biorad) with the following conditions: 15min at 50°C, 2min at 95°C, then 45 times 15sec at 95°C and 40sec at 60°C, data collection occurring during this last step.

Article Title: Pathogen Associated Molecular Pattern (PAMP)-Triggered Immunity Is Compromised under C-Limited Growth
Article Snippet: Total RNA was isolated using RNeasy Plant Mini kit (Qiagen) and the cDNA was synthesized from 2 μg total RNA using oligo(dT) primer and SuperScript III reverse transcriptase (Invitrogen). qRT-PCR was carried out using CFX96 (Bio-Rad) and Quantimix SYBR kit (PhileKorea Technology).

Article Title: Cloning, Expression Profiling and Functional Analysis of CnHMGS, a Gene Encoding 3-hydroxy-3-Methylglutaryl Coenzyme A Synthase from Chamaemelum nobile
Article Snippet: A 1 μg aliquot of the total RNA was used as the template for qRT-PCR. qRT-PCR was performed using a Bio-Rad Mini OpticonTM Real-time PCR Mini Cycler (BioRad, Hercules, CA, USA) with SYBR Premix Ex Taq™ II Kit (Dalian TaKaRa) according to the method of Xu et al. [ ].

Article Title: Repurposing of Tranilast for Potential Neuropathic Pain Treatment by Inhibition of Sepiapterin Reductase in the BH4 Pathway
Article Snippet: Reverse transcription of 500 ng of RNA samples was performed with a Verso cDNA synthesis kit (Thermo Scientific Waltham, USA) according to the manufacturer’s instructions, diluting the product to 1:10 in molecular grade RNA/DNA-free H2 O and stored at −20 °C for use in qRT-PCR. qRT-PCR was performed on a CFX96 RT instrument (Bio-Rad Laboratories, Hercules, US), using triplicates of the samples in a 10 μL reaction containing 5 μL of diluted cDNA (further 1:5 dilution of the stored cDNA synthesis product), 300 nM of the forward and reverse primers and 3 μL of iTaq Universal SYBR Green supermix (Bio-Rad Laboratories, Hercules, US).

Article Title: Bezafibrate Upregulates Mitochondrial Biogenesis and Influence Neural Differentiation of Human-Induced Pluripotent Stem Cells
Article Snippet: qRT-PCR For qRT-PCR, 10 ng of cDNA was loaded with 0.25 μM of forward and reverse primers; 12.5 μL of iTaq™ Universal SYBR® Green Supermix (Bio-rad) onto a 96-well plate for LightCycler® 96 (Roche Diagnostics GmbH) in the following steps: initial denaturation step at 95 °C for 3 min, 45 cycles of denaturation at 95 °C for 10s, and annealing/extension at 58 °C for 1 min.

Article Title: A Novel Sterol Regulatory Element-Binding Protein Gene (sreA) Identified in Penicilliumdigitatum Is Required for Prochloraz Resistance, Full Virulence and erg11 (cyp51) Regulation
Article Snippet: First-strand cDNA was prepared using All-in-one First strand cDNA Synthesis Kit (Genecopoeia, Guangzhou, China) following the manufacturer’s protocol. qRT-PCR was performed using a BIO-RAD CFX96 q-PCR system with SYBR Green I fluorescent dye detection.

Article Title: FXR antagonism of NSAIDs contributes to drug-induced liver injury identified by systems pharmacology approach
Article Snippet: The qRT-PCR was performed using Bio-Rad CFX96TM real time PCR system according to the manufacturer's instruction with specific primers for the FXR target gene SHP (forward 5′-CCCAAGATGCTGTGACCTTT-3′, reverse 5′-CCAGAAGGACTCCAGACAGC-3′), CYP7A1 (forward 5′- GAGAAGGCAAACGGGTGAAC-3′, reverse 5′- GCACAACACCTTATGGTATGACA-3′) for detection of the transcripts.

RNA Sequencing Assay:

Article Title: RNA-seq analysis reveals the role of red light in resistance against Pseudomonas syringae pv. tomato DC3000 in tomato plants
Article Snippet: .. Verification of RNA-seq results by real-time PCR (qRT-PCR) To synthesis cDNA, total RNA from diluted stocks of the same RNA that was subjected to RNA-seq was used in each reverse transcription reaction using the ReverTra Ace qRT-PCR Kit (Toyobo, Japan). qRT-PCR was performed using SYBR-Green chemistry and the iCycler iQ™ Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) [ ]. ..

Quantitative RT-PCR:

Article Title: RNA-seq analysis reveals the role of red light in resistance against Pseudomonas syringae pv. tomato DC3000 in tomato plants
Article Snippet: .. Verification of RNA-seq results by real-time PCR (qRT-PCR) To synthesis cDNA, total RNA from diluted stocks of the same RNA that was subjected to RNA-seq was used in each reverse transcription reaction using the ReverTra Ace qRT-PCR Kit (Toyobo, Japan). qRT-PCR was performed using SYBR-Green chemistry and the iCycler iQ™ Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) [ ]. ..

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    Bio-Rad qrt pcr analysis
    Validation of RNA-seq results by <t>qRT-PCR.</t> Left and right y-axes indicate FPKM values from RNA-seq (blue bar) and relative transcript abundance from qRT-PCR (orange bar). D and S on the x-axis represent Daewonkong and SS0903-2B-21-1-2, respectively. Bars indicate means and standard deviation of three biological replicates. Asterisk above each bar indicates statistical difference between genotypes, as determined by Student’s t-test ( p
    Qrt Pcr Analysis, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 498 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr analysis/product/Bio-Rad
    Average 92 stars, based on 498 article reviews
    Price from $9.99 to $1999.99
    qrt pcr analysis - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    90
    Bio-Rad ddh2 o qrt pcr
    <t>qRT-PCR</t> results of expression of the JH related genes. The expression patterns of 8 genes related to JH in B. mori PG during development in whole fifth larval instar. The relative expression levels were normalized to the Bmrp49 levels. The values are the mean ± SEM (n = 3) of three repeat experiments using qRT-PCR.
    Ddh2 O Qrt Pcr, supplied by Bio-Rad, 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/ddh2 o qrt pcr/product/Bio-Rad
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ddh2 o qrt pcr - by Bioz Stars, 2021-01
    90/100 stars
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    88
    Bio-Rad qrt pcr assay optimization amplification
    In silico dissection approach developing a linear regression model to identify stage-specific gene expression profiles within bulk parasite population gene expression. (A) Definition of physiologically relevant stage categories within P. falciparum development for which we will identify stage-specific expression signatures. Stages are as follows: R: asexual ring, T: asexual trophozoite and schizont, YG: young gametocyte ring and stage I, DG: developing gametocyte stages II, III, and IV, IG: all immature gametocytes (YG+DG), MG: mature gametocyte stage V, and U: unexpected profile not captured by our defined stages. (B) Linear regression model for the deconvolution of bulk gene expression data from mixed stage samples. Terms are as follows: y g : total expression of gene g, β g,s : expression of gene g attributed to stage s, X s : proportion of the sample that is stage s. (C) Marker Selection. Filters used to narrow down gene sets to our set of sentinel markers for field-applicable <t>qRT-PCR</t> assay. As we chose markers for ring and trophozoite/schizont stages a priori based on published stage-specific gene expression data for asexual development [4] , [5] , [46] , we used this selection method to identify markers for the remaining gametocyte stage categories. (D) Overall stage prediction schematic.
    Qrt Pcr Assay Optimization Amplification, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr assay optimization amplification/product/Bio-Rad
    Average 88 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    qrt pcr assay optimization amplification - by Bioz Stars, 2021-01
    88/100 stars
      Buy from Supplier

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    Validation of RNA-seq results by qRT-PCR. Left and right y-axes indicate FPKM values from RNA-seq (blue bar) and relative transcript abundance from qRT-PCR (orange bar). D and S on the x-axis represent Daewonkong and SS0903-2B-21-1-2, respectively. Bars indicate means and standard deviation of three biological replicates. Asterisk above each bar indicates statistical difference between genotypes, as determined by Student’s t-test ( p

    Journal: Scientific Reports

    Article Title: Comprehensive RNA sequencing and co-expression network analysis to complete the biosynthetic pathway of coumestrol, a phytoestrogen

    doi: 10.1038/s41598-018-38219-6

    Figure Lengend Snippet: Validation of RNA-seq results by qRT-PCR. Left and right y-axes indicate FPKM values from RNA-seq (blue bar) and relative transcript abundance from qRT-PCR (orange bar). D and S on the x-axis represent Daewonkong and SS0903-2B-21-1-2, respectively. Bars indicate means and standard deviation of three biological replicates. Asterisk above each bar indicates statistical difference between genotypes, as determined by Student’s t-test ( p

    Article Snippet: qRT-PCR validation of DEGs Gene-specific primers for qRT-PCR analysis were designed based on the nucleotide sequences of selected DEGs using Primer3 ( http://primer3plus.com/ ) (Supplementary Table ). cDNA was synthesized using an iScript™ cDNA Synthesis Kit (Cat. 170-8891; Bio-Rad, Hercules, CA, USA). qRT-PCR was conducted using an iQ™ SYBR Green Supermix kit (Cat. 170-8882; Bio-Rad) on a LightCycler® 480 (Roche Diagnostics, Laval, QC, Canada).

    Techniques: RNA Sequencing Assay, Quantitative RT-PCR, Standard Deviation

    Genetic organization of flhBAFG and fliA genes, and flhF gene expression at 28°C. A . Genetic organization and transcriptional units of flhBAFG and fliA , with arrows showing the direction of transcription. Recognition sites for Eco RI and Stu I are indicated by E and S, respectively. Vertical bars on the map denote the positions and orientation of the Tn 3 - gusA insertion. The two lines with arrowheads beneath the restriction enzyme map indicate the direction and extent of transcription. cDNA synthesized by reverse transcriptase with primers RT1 and RT2 was analyzed by PCR. Thick bars indicate the six expected PCR products, which were verified by electrophoresis on 1.5% agarose gel. B . Photographs of swim assay plates showing the swimming motility of the BGR1 mutant fliA ::Tn 3 - gusA45 and complementation with pBGFA carrying the fliA gene at 28°C and 37°C. C . Expression levels of the flhF gene in the wild-type BGR1, the tofI mutant BGS2, and the qsmR mutant BGS9, and the flhC mutant BGF41 at 28°C, based on qRT-PCR analysis. Vertical lines indicate the standard deviations of three independent experiments.

    Journal: PLoS ONE

    Article Title: Quorum Sensing Controls Flagellar Morphogenesis in Burkholderia glumae

    doi: 10.1371/journal.pone.0084831

    Figure Lengend Snippet: Genetic organization of flhBAFG and fliA genes, and flhF gene expression at 28°C. A . Genetic organization and transcriptional units of flhBAFG and fliA , with arrows showing the direction of transcription. Recognition sites for Eco RI and Stu I are indicated by E and S, respectively. Vertical bars on the map denote the positions and orientation of the Tn 3 - gusA insertion. The two lines with arrowheads beneath the restriction enzyme map indicate the direction and extent of transcription. cDNA synthesized by reverse transcriptase with primers RT1 and RT2 was analyzed by PCR. Thick bars indicate the six expected PCR products, which were verified by electrophoresis on 1.5% agarose gel. B . Photographs of swim assay plates showing the swimming motility of the BGR1 mutant fliA ::Tn 3 - gusA45 and complementation with pBGFA carrying the fliA gene at 28°C and 37°C. C . Expression levels of the flhF gene in the wild-type BGR1, the tofI mutant BGS2, and the qsmR mutant BGS9, and the flhC mutant BGF41 at 28°C, based on qRT-PCR analysis. Vertical lines indicate the standard deviations of three independent experiments.

    Article Snippet: For qRT-PCR analysis, the primers of flhC-cDNA, flhF-cDNA, flhC-qRTF, flhC-qRTR, flhF-qRTF, and flhF-qRTR were used ( ). qRT-PCR analysis was performed according to the manufacturer's instructions except using SsoFast™ EvaGreen Supermix (Bio-Rad) with 25 ng of cDNA as template. qRT-PCR was performed using a thermal cycler (Model C1000™; Bio-Rad) under the following conditions: 98°C for 2 min, followed by 35 cycles at 98°C for 20 s, 65°C for 30 s, and 72°C for 30 s. The 16S rRNA gene was used for data normalization.

    Techniques: Expressing, Synthesized, Polymerase Chain Reaction, Electrophoresis, Agarose Gel Electrophoresis, Mutagenesis, Quantitative RT-PCR

    Expression of flhC and other flagellar genes in the QS mutants at 28°C and 37°C. A . Expression levels of flhC in the wild-type BGR1, the tofI mutant BGS2, and the qsmR mutant BGS9 at 28°C, based on qRT-PCR analysis. Vertical lines indicate the standard deviations of three independent experiments. B . Expression levels of fliC and flgK genes were assessed in the wild-type BGR1, the tofI mutant BGS2, and the qsmR mutant BGS9 at 28°C and 37°C. BGF49, BGR1 fliC ::Tn 3 - gusA49 ; S2F49, BGS2 fliC ::Tn 3 - gusA49 ; S9F49, BGS9 fliC ::Tn 3 - gusA49 ; BGF28, BGR1 flgK ::Tn 3 - gusA28 ; S2F28, BGS2 flgK ::Tn 3 - gusA28 ; S9F28, BGS9 flgK ::Tn 3 - gusA28 .

    Journal: PLoS ONE

    Article Title: Quorum Sensing Controls Flagellar Morphogenesis in Burkholderia glumae

    doi: 10.1371/journal.pone.0084831

    Figure Lengend Snippet: Expression of flhC and other flagellar genes in the QS mutants at 28°C and 37°C. A . Expression levels of flhC in the wild-type BGR1, the tofI mutant BGS2, and the qsmR mutant BGS9 at 28°C, based on qRT-PCR analysis. Vertical lines indicate the standard deviations of three independent experiments. B . Expression levels of fliC and flgK genes were assessed in the wild-type BGR1, the tofI mutant BGS2, and the qsmR mutant BGS9 at 28°C and 37°C. BGF49, BGR1 fliC ::Tn 3 - gusA49 ; S2F49, BGS2 fliC ::Tn 3 - gusA49 ; S9F49, BGS9 fliC ::Tn 3 - gusA49 ; BGF28, BGR1 flgK ::Tn 3 - gusA28 ; S2F28, BGS2 flgK ::Tn 3 - gusA28 ; S9F28, BGS9 flgK ::Tn 3 - gusA28 .

    Article Snippet: For qRT-PCR analysis, the primers of flhC-cDNA, flhF-cDNA, flhC-qRTF, flhC-qRTR, flhF-qRTF, and flhF-qRTR were used ( ). qRT-PCR analysis was performed according to the manufacturer's instructions except using SsoFast™ EvaGreen Supermix (Bio-Rad) with 25 ng of cDNA as template. qRT-PCR was performed using a thermal cycler (Model C1000™; Bio-Rad) under the following conditions: 98°C for 2 min, followed by 35 cycles at 98°C for 20 s, 65°C for 30 s, and 72°C for 30 s. The 16S rRNA gene was used for data normalization.

    Techniques: Expressing, Mutagenesis, Quantitative RT-PCR

    qRT-PCR results of expression of the JH related genes. The expression patterns of 8 genes related to JH in B. mori PG during development in whole fifth larval instar. The relative expression levels were normalized to the Bmrp49 levels. The values are the mean ± SEM (n = 3) of three repeat experiments using qRT-PCR.

    Journal: Scientific Reports

    Article Title: Transcriptomic analysis of the prothoracic gland from two lepidopteran insects, domesticated silkmoth Bombyx mori and wild silkmoth Antheraea pernyi

    doi: 10.1038/s41598-019-41864-0

    Figure Lengend Snippet: qRT-PCR results of expression of the JH related genes. The expression patterns of 8 genes related to JH in B. mori PG during development in whole fifth larval instar. The relative expression levels were normalized to the Bmrp49 levels. The values are the mean ± SEM (n = 3) of three repeat experiments using qRT-PCR.

    Article Snippet: The total volume of qRT-PCR reactions was 10 µl, containing 3.6 µl of TB Green Premix Ex Taq (TaKaRa), 0.4 µl of specific primers (10 µM), 1 µl of cDNA and 5 µl of ddH2 O. qRT-PCR was performed with a BIO-RAD CFX Connect Real-Time System, and the conditions were as follows: 95 °C for 30 s followed by 40 cycles in 95 °C for 5 s and 60 °C for 30 s. Gene-specific primers used for qRT-PCR analysis are listed in Additional file: Table .

    Techniques: Quantitative RT-PCR, Expressing

    qRT-PCR results of expression of the clock genes. The expression patterns of 12 genes related to clock in B. mori PG during development in whole fifth larval instar. The relative expression levels were normalized to the Bmrp49 levels. The values are the mean ± SEM (n = 3) of three repeat experiments using qRT-PCR.

    Journal: Scientific Reports

    Article Title: Transcriptomic analysis of the prothoracic gland from two lepidopteran insects, domesticated silkmoth Bombyx mori and wild silkmoth Antheraea pernyi

    doi: 10.1038/s41598-019-41864-0

    Figure Lengend Snippet: qRT-PCR results of expression of the clock genes. The expression patterns of 12 genes related to clock in B. mori PG during development in whole fifth larval instar. The relative expression levels were normalized to the Bmrp49 levels. The values are the mean ± SEM (n = 3) of three repeat experiments using qRT-PCR.

    Article Snippet: The total volume of qRT-PCR reactions was 10 µl, containing 3.6 µl of TB Green Premix Ex Taq (TaKaRa), 0.4 µl of specific primers (10 µM), 1 µl of cDNA and 5 µl of ddH2 O. qRT-PCR was performed with a BIO-RAD CFX Connect Real-Time System, and the conditions were as follows: 95 °C for 30 s followed by 40 cycles in 95 °C for 5 s and 60 °C for 30 s. Gene-specific primers used for qRT-PCR analysis are listed in Additional file: Table .

    Techniques: Quantitative RT-PCR, Expressing

    In silico dissection approach developing a linear regression model to identify stage-specific gene expression profiles within bulk parasite population gene expression. (A) Definition of physiologically relevant stage categories within P. falciparum development for which we will identify stage-specific expression signatures. Stages are as follows: R: asexual ring, T: asexual trophozoite and schizont, YG: young gametocyte ring and stage I, DG: developing gametocyte stages II, III, and IV, IG: all immature gametocytes (YG+DG), MG: mature gametocyte stage V, and U: unexpected profile not captured by our defined stages. (B) Linear regression model for the deconvolution of bulk gene expression data from mixed stage samples. Terms are as follows: y g : total expression of gene g, β g,s : expression of gene g attributed to stage s, X s : proportion of the sample that is stage s. (C) Marker Selection. Filters used to narrow down gene sets to our set of sentinel markers for field-applicable qRT-PCR assay. As we chose markers for ring and trophozoite/schizont stages a priori based on published stage-specific gene expression data for asexual development [4] , [5] , [46] , we used this selection method to identify markers for the remaining gametocyte stage categories. (D) Overall stage prediction schematic.

    Journal: PLoS Computational Biology

    Article Title: Inferring Developmental Stage Composition from Gene Expression in Human Malaria

    doi: 10.1371/journal.pcbi.1003392

    Figure Lengend Snippet: In silico dissection approach developing a linear regression model to identify stage-specific gene expression profiles within bulk parasite population gene expression. (A) Definition of physiologically relevant stage categories within P. falciparum development for which we will identify stage-specific expression signatures. Stages are as follows: R: asexual ring, T: asexual trophozoite and schizont, YG: young gametocyte ring and stage I, DG: developing gametocyte stages II, III, and IV, IG: all immature gametocytes (YG+DG), MG: mature gametocyte stage V, and U: unexpected profile not captured by our defined stages. (B) Linear regression model for the deconvolution of bulk gene expression data from mixed stage samples. Terms are as follows: y g : total expression of gene g, β g,s : expression of gene g attributed to stage s, X s : proportion of the sample that is stage s. (C) Marker Selection. Filters used to narrow down gene sets to our set of sentinel markers for field-applicable qRT-PCR assay. As we chose markers for ring and trophozoite/schizont stages a priori based on published stage-specific gene expression data for asexual development [4] , [5] , [46] , we used this selection method to identify markers for the remaining gametocyte stage categories. (D) Overall stage prediction schematic.

    Article Snippet: qRT-PCR assay optimization Amplification of the correct target sequence was confirmed by gel electrophoresis and melt curve analysis using SYBR Green (BioRad).

    Techniques: In Silico, Dissection, Expressing, Marker, Selection, Quantitative RT-PCR

    qRT-PCR assay optimization. (A) We collected and analyzed a range of in vitro time points with varying contributions of asexual and sexual stages, from both gametocyte-producing and non-producing lines of 3D7. Absolute number of parasites stages that went into each qRT-PCR reaction well is plotted. (B) Relative qRT-PCR-based gene expression of stage-specific markers for R, T, IG and MG are shown for time points corresponding vertically to those in part A. (C) Inferred proportion of each stage in the total parasite load (model predictions) are shown corresponding vertically to the time points in A and B, plotted as a percentage of total parasites in that sample. (D) In vivo peripheral blood samples from severe malaria patients in Blantyre, Malawi were collected and analyzed. Absolute numbers of parasites stages per µL of blood, as determined by microscopy, are plotted. (E) Relative qRT-PCR-based gene expression of stage-specific markers for T, IG and MG (normalized to SBP1 ) is shown for time points corresponding vertically to those in part D. (F) Inferred proportion of each stage (model predictions) are shown corresponding vertically to the time points in D and E. Stars indicate subjects in which gametocytes were observed by highly sensitive thick smear examination (one or more gametocytes in 100 high power fields).

    Journal: PLoS Computational Biology

    Article Title: Inferring Developmental Stage Composition from Gene Expression in Human Malaria

    doi: 10.1371/journal.pcbi.1003392

    Figure Lengend Snippet: qRT-PCR assay optimization. (A) We collected and analyzed a range of in vitro time points with varying contributions of asexual and sexual stages, from both gametocyte-producing and non-producing lines of 3D7. Absolute number of parasites stages that went into each qRT-PCR reaction well is plotted. (B) Relative qRT-PCR-based gene expression of stage-specific markers for R, T, IG and MG are shown for time points corresponding vertically to those in part A. (C) Inferred proportion of each stage in the total parasite load (model predictions) are shown corresponding vertically to the time points in A and B, plotted as a percentage of total parasites in that sample. (D) In vivo peripheral blood samples from severe malaria patients in Blantyre, Malawi were collected and analyzed. Absolute numbers of parasites stages per µL of blood, as determined by microscopy, are plotted. (E) Relative qRT-PCR-based gene expression of stage-specific markers for T, IG and MG (normalized to SBP1 ) is shown for time points corresponding vertically to those in part D. (F) Inferred proportion of each stage (model predictions) are shown corresponding vertically to the time points in D and E. Stars indicate subjects in which gametocytes were observed by highly sensitive thick smear examination (one or more gametocytes in 100 high power fields).

    Article Snippet: qRT-PCR assay optimization Amplification of the correct target sequence was confirmed by gel electrophoresis and melt curve analysis using SYBR Green (BioRad).

    Techniques: Quantitative RT-PCR, In Vitro, Expressing, In Vivo, Microscopy