cdna template  (Roche)


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    Structured Review

    Roche cdna template
    Standard curves for <t>RT-qPCR</t> assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali <t>cDNA</t> serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.
    Cdna Template, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 137 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cdna template/product/Roche
    Average 94 stars, based on 137 article reviews
    Price from $9.99 to $1999.99
    cdna template - by Bioz Stars, 2020-09
    94/100 stars

    Images

    1) Product Images from "Study on Interactions between the Major Apple Valsa Canker Pathogen Valsa mali and Its Biocontrol Agent Saccharothrix yanglingensis Hhs.015 Using RT-qPCR"

    Article Title: Study on Interactions between the Major Apple Valsa Canker Pathogen Valsa mali and Its Biocontrol Agent Saccharothrix yanglingensis Hhs.015 Using RT-qPCR

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0162174

    Standard curves for RT-qPCR assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali cDNA serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.
    Figure Legend Snippet: Standard curves for RT-qPCR assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali cDNA serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.

    Techniques Used: Quantitative RT-PCR, Expressing, Generated

    2) Product Images from "Characterization of testis-specific serine/threonine kinase 1-like (TSSK1-like) gene and expression patterns in diploid and triploid Pacific abalone (Haliotis discus hannai; Gastropoda; Mollusca) males"

    Article Title: Characterization of testis-specific serine/threonine kinase 1-like (TSSK1-like) gene and expression patterns in diploid and triploid Pacific abalone (Haliotis discus hannai; Gastropoda; Mollusca) males

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0226022

    Expression analysis of TSSK1-like mRNAs in released sperms in comparison with those in mature testes. (A) Representative end-point RT PCR gel showing the amplified TSSK1-like bands from three sperm and two testis total RNA samples. Negative blank (no template) and no RT reaction [RT (−)] showed no amplification. (B) Quantitative measurement of TSSK1-like mRNAs using RT-qPCR analysis. Five testes samples were obtained each from two different half sibling male groups (Testis set-1 and Testis set-2), while five replicate sperm samples were collected from three independent sperm release induction trials (Sperm set-1, -2 and -3). Relative expression levels were normalized based on the direct quantification of cDNA [ 21 ].
    Figure Legend Snippet: Expression analysis of TSSK1-like mRNAs in released sperms in comparison with those in mature testes. (A) Representative end-point RT PCR gel showing the amplified TSSK1-like bands from three sperm and two testis total RNA samples. Negative blank (no template) and no RT reaction [RT (−)] showed no amplification. (B) Quantitative measurement of TSSK1-like mRNAs using RT-qPCR analysis. Five testes samples were obtained each from two different half sibling male groups (Testis set-1 and Testis set-2), while five replicate sperm samples were collected from three independent sperm release induction trials (Sperm set-1, -2 and -3). Relative expression levels were normalized based on the direct quantification of cDNA [ 21 ].

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Quantitative RT-PCR

    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: Study on Interactions between the Major Apple Valsa Canker Pathogen Valsa mali and Its Biocontrol Agent Saccharothrix yanglingensis Hhs.015 Using RT-qPCR
    Article Snippet: .. RT-qPCR assay Experimental samples were analyzed using a Bio-Rad iQ5, qPCR assay in 25 μL reactions containing 1 μL of cDNA template, 0.4 μL of each primers (0.2 μM), 1 × reaction buffer, 2 2 μM MgCl2 , 0.2 2 μM dNTPs (Roche, Mannheim, Germany), 0.4 U Taq DNA polymerase (Thermo Scientific, Lithuania), 2 μL of cDNA template, and 0.5 mL of 2 × SYBR (Takara, Tokyo, Japan). .. The PCR program was as follows: 95°C for 1 min, 40 cycles of 95°C for 10 s, 60°C for 10 s, and 72°C for 40 s. Finally, dissociation curves were generated by increasing the temperature from 65°C to 95°C.

    Article Title: iTRAQ-based comparative proteomic analysis of Vero cells infected with virulent and CV777 vaccine strain-like strains of porcine epidemic diarrhea virus
    Article Snippet: .. Absolute quantitative real-time PCR was performed in a 10 μL volume containing cDNA template, FastStart Universal Probe Master (ROX) (Roche), 100 μM probe and primer. .. The cycle conditions were 50 °C 2 min, 95 °C 10 min, and 40 cycles of 95 °C 15 s, 56 °C 30 s, and 72 °C 31 s. For the standard curve, serial dilutions of a plasmid constructed by inserting the PEDV M gene into the pMD™18-T vector (TAKARA) were used to quantify the virus genomic copy number.

    Article Title: Anti‐hyperlipidaemic effects of synthetic analogues of nordihydroguaiaretic acid in dyslipidaemic rats
    Article Snippet: .. Real‐time PCR was performed in a final volume of 10 μL containing 50 ng of cDNA template and specific sets of forward and reverse primers using a FastStart Universal SYBR Green Master PCR Kit (Roche Applied Science, Indianapolis, IN) and an ABI Prism 7700 system (Applied Biosystems® Life Technologies) according to the manufacturer's protocols. .. The oligonucleotide primers and their sequences used in RT‐PCR are shown in Table .

    Article Title: Characterization of testis-specific serine/threonine kinase 1-like (TSSK1-like) gene and expression patterns in diploid and triploid Pacific abalone (Haliotis discus hannai; Gastropoda; Mollusca) males
    Article Snippet: .. Two μL of the diluted cDNA template was subjected to quantitative PCR amplification using the LightCycler 480 SYBR Green I Master mix and LightCycler 480 Real-Time PCR System (Roche Applied Science, Penzberg, Germany). .. An H . discus hannai RPL5 was also amplified as a normalization control for TSSK1-like expression levels [ , ].

    Article Title: Accessory proteins 8b and 8ab of severe acute respiratory syndrome coronavirus suppress the interferon signaling pathway by mediating ubiquitin-dependent rapid degradation of interferon regulatory factor 3.
    Article Snippet: .. Briefly, a 20 µl PCR reaction containing cDNA template, the respective primers and LightCycler Fast Start SYBR Green I DNA Mastermix (Roche) was prepared and subjected to a qPCR program using the LightCycler (Roche). .. PCR cycling conditions comprised of an initial denaturation step at 94°C for 10 min followed by an amplification program for 40 cycles of 30 s at 95°C, 10 s at 55°C, and 20 s at 72°C with fluorescence acquisition at the end of each extension.

    Amplification:

    Article Title: NUF2 is a valuable prognostic biomarker to predict early recurrence of hepatocellular carcinoma after surgical resection.
    Article Snippet: .. Early tumor recurrence after curative surgical resection poses a great challenge to the clinical management of hepatocellular carcinoma (HCC). .. Early tumor recurrence after curative surgical resection poses a great challenge to the clinical management of hepatocellular carcinoma (HCC).

    Article Title: Characterization of testis-specific serine/threonine kinase 1-like (TSSK1-like) gene and expression patterns in diploid and triploid Pacific abalone (Haliotis discus hannai; Gastropoda; Mollusca) males
    Article Snippet: .. Two μL of the diluted cDNA template was subjected to quantitative PCR amplification using the LightCycler 480 SYBR Green I Master mix and LightCycler 480 Real-Time PCR System (Roche Applied Science, Penzberg, Germany). .. An H . discus hannai RPL5 was also amplified as a normalization control for TSSK1-like expression levels [ , ].

    Article Title: KH-type splicing regulatory protein controls colorectal cancer cell growth and modulates the tumor microenvironment
    Article Snippet: .. 1 μg of RNA was reverse-transcribed into cDNA using the RT Omniscript cDNA kit (Qiagen), and 20 ng of cDNA template was amplified on a LightCycler-480 (Roche, Indianapolis, IL, USA) using a FAST SYBR Green Master Mix (Invitrogen) under the following conditions: pre-incubation step at 95 °C for 5 min was followed by 45 cycles of denaturation at 95 °C for 10 sec, annealing at 60 °C for 10 sec, and elongation at 72 °C for 10 sec. GAPDH was used as the housekeeping gene. .. The mRNA expression levels of all samples were normalised to the housekeeping gene, and the delta/delta Ct method was used to calculate relative expression (treatment vs. control).

    Quantitative RT-PCR:

    Article Title: Study on Interactions between the Major Apple Valsa Canker Pathogen Valsa mali and Its Biocontrol Agent Saccharothrix yanglingensis Hhs.015 Using RT-qPCR
    Article Snippet: .. RT-qPCR assay Experimental samples were analyzed using a Bio-Rad iQ5, qPCR assay in 25 μL reactions containing 1 μL of cDNA template, 0.4 μL of each primers (0.2 μM), 1 × reaction buffer, 2 2 μM MgCl2 , 0.2 2 μM dNTPs (Roche, Mannheim, Germany), 0.4 U Taq DNA polymerase (Thermo Scientific, Lithuania), 2 μL of cDNA template, and 0.5 mL of 2 × SYBR (Takara, Tokyo, Japan). .. The PCR program was as follows: 95°C for 1 min, 40 cycles of 95°C for 10 s, 60°C for 10 s, and 72°C for 40 s. Finally, dissociation curves were generated by increasing the temperature from 65°C to 95°C.

    SYBR Green Assay:

    Article Title: Anti‐hyperlipidaemic effects of synthetic analogues of nordihydroguaiaretic acid in dyslipidaemic rats
    Article Snippet: .. Real‐time PCR was performed in a final volume of 10 μL containing 50 ng of cDNA template and specific sets of forward and reverse primers using a FastStart Universal SYBR Green Master PCR Kit (Roche Applied Science, Indianapolis, IN) and an ABI Prism 7700 system (Applied Biosystems® Life Technologies) according to the manufacturer's protocols. .. The oligonucleotide primers and their sequences used in RT‐PCR are shown in Table .

    Article Title: Characterization of testis-specific serine/threonine kinase 1-like (TSSK1-like) gene and expression patterns in diploid and triploid Pacific abalone (Haliotis discus hannai; Gastropoda; Mollusca) males
    Article Snippet: .. Two μL of the diluted cDNA template was subjected to quantitative PCR amplification using the LightCycler 480 SYBR Green I Master mix and LightCycler 480 Real-Time PCR System (Roche Applied Science, Penzberg, Germany). .. An H . discus hannai RPL5 was also amplified as a normalization control for TSSK1-like expression levels [ , ].

    Article Title: Accessory proteins 8b and 8ab of severe acute respiratory syndrome coronavirus suppress the interferon signaling pathway by mediating ubiquitin-dependent rapid degradation of interferon regulatory factor 3.
    Article Snippet: .. Briefly, a 20 µl PCR reaction containing cDNA template, the respective primers and LightCycler Fast Start SYBR Green I DNA Mastermix (Roche) was prepared and subjected to a qPCR program using the LightCycler (Roche). .. PCR cycling conditions comprised of an initial denaturation step at 94°C for 10 min followed by an amplification program for 40 cycles of 30 s at 95°C, 10 s at 55°C, and 20 s at 72°C with fluorescence acquisition at the end of each extension.

    Article Title: KH-type splicing regulatory protein controls colorectal cancer cell growth and modulates the tumor microenvironment
    Article Snippet: .. 1 μg of RNA was reverse-transcribed into cDNA using the RT Omniscript cDNA kit (Qiagen), and 20 ng of cDNA template was amplified on a LightCycler-480 (Roche, Indianapolis, IL, USA) using a FAST SYBR Green Master Mix (Invitrogen) under the following conditions: pre-incubation step at 95 °C for 5 min was followed by 45 cycles of denaturation at 95 °C for 10 sec, annealing at 60 °C for 10 sec, and elongation at 72 °C for 10 sec. GAPDH was used as the housekeeping gene. .. The mRNA expression levels of all samples were normalised to the housekeeping gene, and the delta/delta Ct method was used to calculate relative expression (treatment vs. control).

    Article Title: Quantitative proteomic analysis reveals that transmissible gastroenteritis virus activates the JAK-STAT1 signaling pathway.
    Article Snippet: .. Transmissible gastroenteritis virus (TGEV), a porcine enteropathogenic coronavirus, causes lethal watery diarrhea and severe dehydration in piglets. .. Transmissible gastroenteritis virus (TGEV), a porcine enteropathogenic coronavirus, causes lethal watery diarrhea and severe dehydration in piglets.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Quantitative proteomic analysis reveals that transmissible gastroenteritis virus activates the JAK-STAT1 signaling pathway.
    Article Snippet: .. Transmissible gastroenteritis virus (TGEV), a porcine enteropathogenic coronavirus, causes lethal watery diarrhea and severe dehydration in piglets. .. Transmissible gastroenteritis virus (TGEV), a porcine enteropathogenic coronavirus, causes lethal watery diarrhea and severe dehydration in piglets.

    Polymerase Chain Reaction:

    Article Title: NUF2 is a valuable prognostic biomarker to predict early recurrence of hepatocellular carcinoma after surgical resection.
    Article Snippet: .. Early tumor recurrence after curative surgical resection poses a great challenge to the clinical management of hepatocellular carcinoma (HCC). .. Early tumor recurrence after curative surgical resection poses a great challenge to the clinical management of hepatocellular carcinoma (HCC).

    Article Title: Anti‐hyperlipidaemic effects of synthetic analogues of nordihydroguaiaretic acid in dyslipidaemic rats
    Article Snippet: .. Real‐time PCR was performed in a final volume of 10 μL containing 50 ng of cDNA template and specific sets of forward and reverse primers using a FastStart Universal SYBR Green Master PCR Kit (Roche Applied Science, Indianapolis, IN) and an ABI Prism 7700 system (Applied Biosystems® Life Technologies) according to the manufacturer's protocols. .. The oligonucleotide primers and their sequences used in RT‐PCR are shown in Table .

    Article Title: Accessory proteins 8b and 8ab of severe acute respiratory syndrome coronavirus suppress the interferon signaling pathway by mediating ubiquitin-dependent rapid degradation of interferon regulatory factor 3.
    Article Snippet: .. Briefly, a 20 µl PCR reaction containing cDNA template, the respective primers and LightCycler Fast Start SYBR Green I DNA Mastermix (Roche) was prepared and subjected to a qPCR program using the LightCycler (Roche). .. PCR cycling conditions comprised of an initial denaturation step at 94°C for 10 min followed by an amplification program for 40 cycles of 30 s at 95°C, 10 s at 55°C, and 20 s at 72°C with fluorescence acquisition at the end of each extension.

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  • 94
    Roche cdna template
    Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into <t>cDNA.</t> The <t>qPCR</t> was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values
    Cdna Template, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 198 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cdna template/product/Roche
    Average 94 stars, based on 198 article reviews
    Price from $9.99 to $1999.99
    cdna template - by Bioz Stars, 2020-09
    94/100 stars
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    Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into cDNA. The qPCR was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values

    Journal: Scientific Reports

    Article Title: Small secreted proteins from the necrotrophic conifer pathogen Heterobasidion annosum s.l. (HaSSPs) induce cell death in Nicotiana benthamiana

    doi: 10.1038/s41598-017-08010-0

    Figure Lengend Snippet: Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into cDNA. The qPCR was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values

    Article Snippet: The qPCR reaction mixture was assembled with 5.5 µl cDNA template (1:15 dilution), 1 µl forward primer, 1 µl reverse primer, and 7.5 µl 2X LightCycler 480 SYBR Green I Master (Roche, Finland).

    Techniques: Expressing, Plasmid Preparation, Incubation, Real-time Polymerase Chain Reaction, Activation Assay, Standard Deviation, Two Tailed Test

    KGFR and KGF in pancreatic cancer cell lines. A: RT-PCR analysis of KGFR and KGF mRNA in pancreatic cancer cell lines. Total RNA was extracted from PANC-1; MIA PaCa-2; KLM-1; Capan-1; and PK-1, -8, -9, and -59 cells and used to perform cDNA synthesis and PCR analysis. KGFR mRNA (150 bp) was detected in all cell lines (top). KGF mRNA (163-bp) was detected in all pancreatic cancer cell lines, except for MIA PaCa-2 cells (middle). β-Actin mRNA (163 bp) served as a loading control (bottom). B and C: Q-PCR analysis of KGFR and KGF mRNA in pancreatic cancer cell lines. KGFR mRNA was expressed at variable levels in all pancreatic cancer cell lines. KGF mRNA was not detected in MIA PaCa-2 cells and was expressed at variable levels in the other pancreatic cancer cell lines. Results are expressed as KGFR/β-actin and KGF/β-actin. Gene expression measurements were performed in triplicate. Bars represent the mean ± SE. D: Western blot analysis of KGFR in pancreatic cancer cell lines. A band corresponding to the 105-kd KGFR protein was detected in all pancreatic cancer cell lines (top). β-Actin served as a loading control (bottom). E: Immunofluorescent analysis of KGFR expression in MIA PaCa-2 cells. KGFR immunoreactivity was detected in the cytoplasm and/or on the membrane of cancer cells. In some cases, a strong KGFR signal was detected at the cell membrane ( arrowheads ). Bar = 50 μm.

    Journal: The American Journal of Pathology

    Article Title: Enhanced Expression of Keratinocyte Growth Factor and Its Receptor Correlates with Venous Invasion in Pancreatic Cancer

    doi: 10.2353/ajpath.2007.060935

    Figure Lengend Snippet: KGFR and KGF in pancreatic cancer cell lines. A: RT-PCR analysis of KGFR and KGF mRNA in pancreatic cancer cell lines. Total RNA was extracted from PANC-1; MIA PaCa-2; KLM-1; Capan-1; and PK-1, -8, -9, and -59 cells and used to perform cDNA synthesis and PCR analysis. KGFR mRNA (150 bp) was detected in all cell lines (top). KGF mRNA (163-bp) was detected in all pancreatic cancer cell lines, except for MIA PaCa-2 cells (middle). β-Actin mRNA (163 bp) served as a loading control (bottom). B and C: Q-PCR analysis of KGFR and KGF mRNA in pancreatic cancer cell lines. KGFR mRNA was expressed at variable levels in all pancreatic cancer cell lines. KGF mRNA was not detected in MIA PaCa-2 cells and was expressed at variable levels in the other pancreatic cancer cell lines. Results are expressed as KGFR/β-actin and KGF/β-actin. Gene expression measurements were performed in triplicate. Bars represent the mean ± SE. D: Western blot analysis of KGFR in pancreatic cancer cell lines. A band corresponding to the 105-kd KGFR protein was detected in all pancreatic cancer cell lines (top). β-Actin served as a loading control (bottom). E: Immunofluorescent analysis of KGFR expression in MIA PaCa-2 cells. KGFR immunoreactivity was detected in the cytoplasm and/or on the membrane of cancer cells. In some cases, a strong KGFR signal was detected at the cell membrane ( arrowheads ). Bar = 50 μm.

    Article Snippet: PCR reaction mixture containing 2 μl of template cDNA, 3 mmol/L MgCl2 , and 0.5 μmol/L of primers, and LightCycler-FastStart DNA Master SYBR Green I mix was applied into a capillary tube (Roche).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Expressing, Western Blot

    Expression of AgCad1 in A. gambiae ) gut cDNA with AgCad1 primers showing

    Journal: Biochemistry

    Article Title: Anopheles gambiae Cadherin AgCad1 Binds the Cry4Ba Toxin of Bacillus thuringiensis israelensis and a Fragment of AgCad1 Synergizes Toxicity †

    doi: 10.1021/bi7023578

    Figure Lengend Snippet: Expression of AgCad1 in A. gambiae ) gut cDNA with AgCad1 primers showing

    Article Snippet: Oligonucleotide primers AgCad1/F-BamH and AgCad1/R-Sac were used to amplify the 3′ end of the AgCad1 coding region using the cDNA template, using an Expand Long Template PCR System (Roche Applied Science, Indianapolis, IN) with 30 cycles of 94 °C for 2 min and 68 °C for 2 min.

    Techniques: Expressing

    Standard curves for RT-qPCR assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali cDNA serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.

    Journal: PLoS ONE

    Article Title: Study on Interactions between the Major Apple Valsa Canker Pathogen Valsa mali and Its Biocontrol Agent Saccharothrix yanglingensis Hhs.015 Using RT-qPCR

    doi: 10.1371/journal.pone.0162174

    Figure Lengend Snippet: Standard curves for RT-qPCR assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali cDNA serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.

    Article Snippet: RT-qPCR assay Experimental samples were analyzed using a Bio-Rad iQ5, qPCR assay in 25 μL reactions containing 1 μL of cDNA template, 0.4 μL of each primers (0.2 μM), 1 × reaction buffer, 2 2 μM MgCl2 , 0.2 2 μM dNTPs (Roche, Mannheim, Germany), 0.4 U Taq DNA polymerase (Thermo Scientific, Lithuania), 2 μL of cDNA template, and 0.5 mL of 2 × SYBR (Takara, Tokyo, Japan).

    Techniques: Quantitative RT-PCR, Expressing, Generated