first-strand complementary dna reaction Search Results


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  • 99
    New England Biolabs first strand complementary dna reaction
    qPCR analyses of candidate circRNA expression. Cells were treated with DMSO (control) and cisplatin as explained in Materials and Methods. RNAse R-treated RNA samples were used to prepare cDNAs using <t>ProtoScript</t> ® first strand <t>cDNA</t> synthesis kit (NEB, United States) with random primers. GoTaq q-PCR Master Mix (Promega, United States) was used for qPCR analyses. Data collected in three biological replicates were normalized against DMSO control and GAPDH. ∗∗ and ∗∗∗ denote p ≤ 0.01 and p ≤ 0.001, respectively. (A) HeLa cells; (B) MCF and Jurkat cells.
    First Strand Complementary Dna Reaction, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher first strand cdna synthesis reaction
    Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, <t>Vpr,</t> Rev, and Nef as marked on the top. Panel D. <t>cDNA</t> obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.
    First Strand Cdna Synthesis Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2845 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher complementary dna cdna
    Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, <t>Vpr,</t> Rev, and Nef as marked on the top. Panel D. <t>cDNA</t> obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.
    Complementary Dna Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 20305 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bio-Rad strand complementary dna cdna
    Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, <t>Vpr,</t> Rev, and Nef as marked on the top. Panel D. <t>cDNA</t> obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.
    Strand Complementary Dna Cdna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Quanta Biosciences first strand cdna reaction
    Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, <t>Vpr,</t> Rev, and Nef as marked on the top. Panel D. <t>cDNA</t> obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.
    First Strand Cdna Reaction, supplied by Quanta Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cdna  (Roche)
    99
    Roche cdna
    Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, <t>Vpr,</t> Rev, and Nef as marked on the top. Panel D. <t>cDNA</t> obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.
    Cdna, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 3381 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    TaKaRa complementary dna cdna strand
    Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, <t>Vpr,</t> Rev, and Nef as marked on the top. Panel D. <t>cDNA</t> obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.
    Complementary Dna Cdna Strand, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Promega strand cdna synthesis kit
    Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, <t>Vpr,</t> Rev, and Nef as marked on the top. Panel D. <t>cDNA</t> obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.
    Strand Cdna Synthesis Kit, supplied by Promega, used in various techniques. Bioz Stars score: 95/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa first strand complementary dna cdna
    qPCR assay. (A) Gel electrophoresis analysis of the target and reference gene PCR products. Lanes 1 and 6, 500-bp molecular marker; 2, LRIG2; 3, NTC for LRIG2; 4, GAPDH; and 5, NTC for GAPDH. (B) Melting curve analysis of the target and reference gene PCR products. The SYBR Green qPCR reactions of the (a) LRIG2 and (b) GAPDH genes were performed using a normal sample of the complementary <t>DNA</t> <t>(cDNA).</t> Each peak corresponds to a unique PCR product. (c) NTC reactions showed no PCR product. LRIG2, leucine-rich repeats and immunoglobulin-like domains 2; NTC, non-template negative control; qPCR, quantitative polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; −dI/dT, negative first derivative of the melting curves as a function of temperature.
    First Strand Complementary Dna Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 747 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Bio-Rad first strand complementary dna cdna
    qPCR assay. (A) Gel electrophoresis analysis of the target and reference gene PCR products. Lanes 1 and 6, 500-bp molecular marker; 2, LRIG2; 3, NTC for LRIG2; 4, GAPDH; and 5, NTC for GAPDH. (B) Melting curve analysis of the target and reference gene PCR products. The SYBR Green qPCR reactions of the (a) LRIG2 and (b) GAPDH genes were performed using a normal sample of the complementary <t>DNA</t> <t>(cDNA).</t> Each peak corresponds to a unique PCR product. (c) NTC reactions showed no PCR product. LRIG2, leucine-rich repeats and immunoglobulin-like domains 2; NTC, non-template negative control; qPCR, quantitative polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; −dI/dT, negative first derivative of the melting curves as a function of temperature.
    First Strand Complementary Dna Cdna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 98/100, based on 442 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher cdna synthesis reaction
    qPCR assay. (A) Gel electrophoresis analysis of the target and reference gene PCR products. Lanes 1 and 6, 500-bp molecular marker; 2, LRIG2; 3, NTC for LRIG2; 4, GAPDH; and 5, NTC for GAPDH. (B) Melting curve analysis of the target and reference gene PCR products. The SYBR Green qPCR reactions of the (a) LRIG2 and (b) GAPDH genes were performed using a normal sample of the complementary <t>DNA</t> <t>(cDNA).</t> Each peak corresponds to a unique PCR product. (c) NTC reactions showed no PCR product. LRIG2, leucine-rich repeats and immunoglobulin-like domains 2; NTC, non-template negative control; qPCR, quantitative polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; −dI/dT, negative first derivative of the melting curves as a function of temperature.
    Cdna Synthesis Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 946 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    GE Healthcare first strand cdna synthesis
    qPCR assay. (A) Gel electrophoresis analysis of the target and reference gene PCR products. Lanes 1 and 6, 500-bp molecular marker; 2, LRIG2; 3, NTC for LRIG2; 4, GAPDH; and 5, NTC for GAPDH. (B) Melting curve analysis of the target and reference gene PCR products. The SYBR Green qPCR reactions of the (a) LRIG2 and (b) GAPDH genes were performed using a normal sample of the complementary <t>DNA</t> <t>(cDNA).</t> Each peak corresponds to a unique PCR product. (c) NTC reactions showed no PCR product. LRIG2, leucine-rich repeats and immunoglobulin-like domains 2; NTC, non-template negative control; qPCR, quantitative polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; −dI/dT, negative first derivative of the melting curves as a function of temperature.
    First Strand Cdna Synthesis, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 94/100, based on 522 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Roche transcriptor first strand complementary dna cdna synthesis kit
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    Transcriptor First Strand Complementary Dna Cdna Synthesis Kit, supplied by Roche, used in various techniques. Bioz Stars score: 96/100, based on 908 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    SuperArray Bioscience Corporation first strand complementary dna cdna
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    First Strand Complementary Dna Cdna, supplied by SuperArray Bioscience Corporation, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    OriGene first strand complementary dna cdna
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    First Strand Complementary Dna Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    GE Healthcare cdna first strand synthesis reaction
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    Cdna First Strand Synthesis Reaction, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche first strand complementary dna cdna
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    First Strand Complementary Dna Cdna, supplied by Roche, used in various techniques. Bioz Stars score: 97/100, based on 151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Stratagene first strand complementary dna cdna
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    First Strand Complementary Dna Cdna, supplied by Stratagene, used in various techniques. Bioz Stars score: 86/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Toyobo first strand complementary dna cdna
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    First Strand Complementary Dna Cdna, supplied by Toyobo, used in various techniques. Bioz Stars score: 93/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co strand complementary dna cdna synthesis
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    Strand Complementary Dna Cdna Synthesis, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 90/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATUM complimentary dna
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    Complimentary Dna, supplied by ATUM, used in various techniques. Bioz Stars score: 82/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher strand complementary dna cdna synthesis kit
    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
    Strand Complementary Dna Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
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    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand <t>cDNA,</t> <t>transcriptor</t> first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P
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    Evaluation of subtraction efficiency. A: Reduction of GAPDH <t>cDNA</t> following subtraction in the SH-Ctls sample. <t>PCR</t> was performed on SH-Ctls subtracted and SH unsubtracted samples. GAPDH PCR products (760 pb) were detectable 10 cycles earlier in the unsubtracted sample (18 cycles) than in the subtracted sample (28 cycles). B: Enrichment of LCN2 cDNA following subtraction in the SH-Ctls sample. PCR was performed on SH-Ctls and Ctls-SH subtracted samples as well as SH and Ctls unsubtracted samples. LCN2 PCR products (210 pb) were detected after 20 cycles for both SH unsubtracted and SH-Ctls subtracted samples, the difference in the intensity of the 2 bands indicate the enrichment compare to Ctls unsubtracted and Ctls-SH subtracted samples.
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    Validation of alternative splicing events co-regulated by JMJD6 and U2AF65 through RT-PCR. ( A–J ) First-strand <t>cDNA</t> synthesis was done from <t>RNA</t> samples described in Figure 1B followed by standard PCR analysis (RT-PCR) using primer sets specifically targeting to the upstream and downstream exon relative to the cassette exon (alternatively spliced exon) to detect the expression of both short and long isoforms of representative genes as indicated. The length of the alternatively spliced exon as well as the expected length of the PCR products was shown as indicated. DNA marker (M) was included on the left. Percentage spliced in (PSI) was calculated as the ratio of the density of the long isoform versus that of the sum of the long and short isoforms. The position of the alternatively spliced exon in each gene was as follows: KIAA0515 (NM_013318, exon 30) (A); TCERG1 (BI091338, exon 3) (B); BAT2D1 (AV650960, exon 6) (C); HNRNPH3 (NM_012207, exon 3) (D); UTRN (NM_007124, exon 66) (E); TMEM126B (NM_018480, exon 2) (F); ZDHHC20 (uc001uod.1, exon 13) (G); BRD8 (NM_006696, exon 21) (H); PTBLP (NM_021190, exon 10) (I); NCAM1 (NM_181351, exon 9) (J). F: forward primer; R: reverse primer; bp: base pair.
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    Validation of alternative splicing events co-regulated by JMJD6 and U2AF65 through RT-PCR. ( A–J ) First-strand <t>cDNA</t> synthesis was done from <t>RNA</t> samples described in Figure 1B followed by standard PCR analysis (RT-PCR) using primer sets specifically targeting to the upstream and downstream exon relative to the cassette exon (alternatively spliced exon) to detect the expression of both short and long isoforms of representative genes as indicated. The length of the alternatively spliced exon as well as the expected length of the PCR products was shown as indicated. DNA marker (M) was included on the left. Percentage spliced in (PSI) was calculated as the ratio of the density of the long isoform versus that of the sum of the long and short isoforms. The position of the alternatively spliced exon in each gene was as follows: KIAA0515 (NM_013318, exon 30) (A); TCERG1 (BI091338, exon 3) (B); BAT2D1 (AV650960, exon 6) (C); HNRNPH3 (NM_012207, exon 3) (D); UTRN (NM_007124, exon 66) (E); TMEM126B (NM_018480, exon 2) (F); ZDHHC20 (uc001uod.1, exon 13) (G); BRD8 (NM_006696, exon 21) (H); PTBLP (NM_021190, exon 10) (I); NCAM1 (NM_181351, exon 9) (J). F: forward primer; R: reverse primer; bp: base pair.
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    Adrenomedullin expression by breast cancer cell lines and effects of adrenomedullin on tumor and bone cells. (A) Adrenomedullin ( AM) RNA in human breast cancer cell lines that metastasize to bone. <t>PCR</t> detection of human AM mRNA in from left to right: normal tissues (kidney, prostate and breast), breast cancer cell lines (MCF-7, BT549, MDA-231, MCF-7, MDA-MB-436, ZR-75-1, T47D, MD-MB-361 and BT438) and three additional cancer cell lines that cause osteolytic bone metastases: MDA-MB-435S melanoma cells, PC-3 prostate cancer cells and A549 lung adenocarcinoma cells. <t>cDNA</t> (200 ng) was amplified for 30 cycles. The PCR product is 600 bp. It is unclear if kidney expresses only low amounts of AM mRNA or if the RNA was degraded. (B) Stable overexpression of human AM mRNA in MDA-MB-231 cells. Clones were tested for stable human AM mRNA expression using real-time PCR after 60 days of culture in the absence of antibiotic selection. G8-G31 are MDA-MB-231 control clones, and A15-A51 are MDA-MB-231 clones overexpressing AM. (C) Proliferation in vitro of breast cancer cells. Proliferation of two AM-overexpressing clones (A39, gray triangle; A51, black triangle) and two control clones (G8, gray square; G26, black square) were compared using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; tetrazolium blue dye) assay. Proliferation was compared after 1, 3 and 5 days of growth. OD, Optical density.
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    Image Search Results


    qPCR analyses of candidate circRNA expression. Cells were treated with DMSO (control) and cisplatin as explained in Materials and Methods. RNAse R-treated RNA samples were used to prepare cDNAs using ProtoScript ® first strand cDNA synthesis kit (NEB, United States) with random primers. GoTaq q-PCR Master Mix (Promega, United States) was used for qPCR analyses. Data collected in three biological replicates were normalized against DMSO control and GAPDH. ∗∗ and ∗∗∗ denote p ≤ 0.01 and p ≤ 0.001, respectively. (A) HeLa cells; (B) MCF and Jurkat cells.

    Journal: Frontiers in Genetics

    Article Title: Transcriptomics Analysis of Circular RNAs Differentially Expressed in Apoptotic HeLa Cells

    doi: 10.3389/fgene.2019.00176

    Figure Lengend Snippet: qPCR analyses of candidate circRNA expression. Cells were treated with DMSO (control) and cisplatin as explained in Materials and Methods. RNAse R-treated RNA samples were used to prepare cDNAs using ProtoScript ® first strand cDNA synthesis kit (NEB, United States) with random primers. GoTaq q-PCR Master Mix (Promega, United States) was used for qPCR analyses. Data collected in three biological replicates were normalized against DMSO control and GAPDH. ∗∗ and ∗∗∗ denote p ≤ 0.01 and p ≤ 0.001, respectively. (A) HeLa cells; (B) MCF and Jurkat cells.

    Article Snippet: Subsequently, RNA clean-up was performed with Nucleospin RNA kit (Macherey Nagel, Germany) according to the manufacturer’s instructions. cDNAs were prepared from RNAse R+ and RNAseR- RNA samples with ProtoScript® first strand cDNA synthesis kit (NEB, United States) with random primers in a total volume of 50 ul according to the manufacturer’s instructions.

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Polymerase Chain Reaction

    Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, Vpr, Rev, and Nef as marked on the top. Panel D. cDNA obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.

    Journal: PLoS ONE

    Article Title: Multiplex RT-PCR Amplification of HIV Genes to Create a Completely Autologous DC-Based Immunotherapy for the Treatment of HIV Infection

    doi: 10.1371/journal.pone.0001489

    Figure Lengend Snippet: Successful clade-independent amplification of HIV RNA encoding for antigens from infectious plasma. Panel A: Agarose gel electrophoresis analysis of PCR fragment obtained from three diverse plasma. Amplification from subject plasma infected with Clade B sample. M: 100 bp DNA ladder (Invitrogen). Panel B: Amplification from subject plasma infected with Clade C virus. M: 100 bp DNA ladder (Invitrogen). Panel C: Amplification from subject plasma infected with Clade AG virus. M: AmpliSize DNA ladder (BioRad). Analysis of products obtained after the secondary PCR reaction for Gag, Vpr, Rev, and Nef as marked on the top. Panel D. cDNA obtained in preparative secondary PCR reaction corresponding to Gag, Vpr, Rev, and Nef antigens. M: 100 bp DNA ladder (Invitrogen). The molecular weight of representative DNA bands is indicated on the left. Panel E. RNA corresponding to Gag, Vpr, Rev, and Nef antigens obtained by in vitro transcription using amplified PCR products from subjects plasma. M: molecular weight RNA ladder (Promega), representative marker sizes are indicated on the left. G, V, R, N: in vitro transcribed RNAs for Gag, Vpr, Nef and Nef respectively.

    Article Snippet: First strand cDNA synthesis reaction contained gene-specific primers for either Gag, Vpr or Rev, and oligo dT(20) (Invitrogen) for Nef, 40 units of RNAseOut (Invitrogen), 0.5 mM of each dNTP (Clontech), and Superscript first strand buffer.

    Techniques: Amplification, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Infection, Molecular Weight, In Vitro, Marker

    Gel analysis of F. rubra and E. festucae antisense transcripts. The diagram illustrates primer design for detection of sense and antisense transcripts. The “A” primers were used for strand specific synthesis of cDNA from the RNA sample. The “A” and “B” primers were used for cDNA amplification. cDNAs generated from gene-specific primers for the F. rubra metallothionein (MT) and the E. festucae NC12, antifungal protein (AFP), and subtilisin-like protease were used as templates for PCR amplification.

    Journal: PLoS ONE

    Article Title: SOLiD-SAGE of Endophyte-Infected Red Fescue Reveals Numerous Effects on Host Transcriptome and an Abundance of Highly Expressed Fungal Secreted Proteins

    doi: 10.1371/journal.pone.0053214

    Figure Lengend Snippet: Gel analysis of F. rubra and E. festucae antisense transcripts. The diagram illustrates primer design for detection of sense and antisense transcripts. The “A” primers were used for strand specific synthesis of cDNA from the RNA sample. The “A” and “B” primers were used for cDNA amplification. cDNAs generated from gene-specific primers for the F. rubra metallothionein (MT) and the E. festucae NC12, antifungal protein (AFP), and subtilisin-like protease were used as templates for PCR amplification.

    Article Snippet: First-strand cDNA of 4 µg S1139RC total RNA was synthesized from either 500 ng of oligo(dT)18 primer or 2 picomoles of a strand-specific primer by using SuperScript™ III Reverse Transcriptase (Life Technologies, Carlsbad, CA) according to the manufacturer’s instructions.

    Techniques: Amplification, Generated, Polymerase Chain Reaction

    Dectin-1 silencing. DCs were electroporated with Dectin-1 siRNA or non-silencing siRNA and were cultured for 24 h at 37°C. RNA was isolated from 1 × 10 6 cells and was converted to cDNA. RT-PCR was performed to quantify Dectin-1 downregulation on mRNA level. Dectin-1 mRNA expression level of Dectin-1 silenced DCs was calculated compared to a non-silencing control. Data are represented as mean + SEM of n = 5 independent experiments. A student's t -test was performed and significant differences are marked with an asterisk (*** p

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: First Insights in NK—DC Cross-Talk and the Importance of Soluble Factors During Infection With Aspergillus fumigatus

    doi: 10.3389/fcimb.2018.00288

    Figure Lengend Snippet: Dectin-1 silencing. DCs were electroporated with Dectin-1 siRNA or non-silencing siRNA and were cultured for 24 h at 37°C. RNA was isolated from 1 × 10 6 cells and was converted to cDNA. RT-PCR was performed to quantify Dectin-1 downregulation on mRNA level. Dectin-1 mRNA expression level of Dectin-1 silenced DCs was calculated compared to a non-silencing control. Data are represented as mean + SEM of n = 5 independent experiments. A student's t -test was performed and significant differences are marked with an asterisk (*** p

    Article Snippet: Therefore, RNA was isolated from mock silenced and Dectin-1 silenced DCs by RNeasy Mini kit (Qiagen) before cDNA synthesis (First Strand cDNA Synthesis Kit, K1612, Thermo Fisher Scientific) was performed.

    Techniques: Cell Culture, Isolation, Reverse Transcription Polymerase Chain Reaction, Expressing

    qPCR assay. (A) Gel electrophoresis analysis of the target and reference gene PCR products. Lanes 1 and 6, 500-bp molecular marker; 2, LRIG2; 3, NTC for LRIG2; 4, GAPDH; and 5, NTC for GAPDH. (B) Melting curve analysis of the target and reference gene PCR products. The SYBR Green qPCR reactions of the (a) LRIG2 and (b) GAPDH genes were performed using a normal sample of the complementary DNA (cDNA). Each peak corresponds to a unique PCR product. (c) NTC reactions showed no PCR product. LRIG2, leucine-rich repeats and immunoglobulin-like domains 2; NTC, non-template negative control; qPCR, quantitative polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; −dI/dT, negative first derivative of the melting curves as a function of temperature.

    Journal: Oncology Letters

    Article Title: LRIG2 expression and prognosis in non-small cell lung cancer

    doi: 10.3892/ol.2014.2157

    Figure Lengend Snippet: qPCR assay. (A) Gel electrophoresis analysis of the target and reference gene PCR products. Lanes 1 and 6, 500-bp molecular marker; 2, LRIG2; 3, NTC for LRIG2; 4, GAPDH; and 5, NTC for GAPDH. (B) Melting curve analysis of the target and reference gene PCR products. The SYBR Green qPCR reactions of the (a) LRIG2 and (b) GAPDH genes were performed using a normal sample of the complementary DNA (cDNA). Each peak corresponds to a unique PCR product. (c) NTC reactions showed no PCR product. LRIG2, leucine-rich repeats and immunoglobulin-like domains 2; NTC, non-template negative control; qPCR, quantitative polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; −dI/dT, negative first derivative of the melting curves as a function of temperature.

    Article Snippet: The first-strand complementary DNA (cDNA) was prepared from total RNA using a first-strand PrimeScript™ RT reagent kit with gDNA Eraser (Takara Bio, Inc., Shiga, Japan).

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

    The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand cDNA, transcriptor first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P

    Journal: Cell Death Discovery

    Article Title: TRAIL DR5-CTSB crosstalk participates in breast cancer autophagy initiated by SAHA

    doi: 10.1038/cddiscovery.2017.52

    Figure Lengend Snippet: The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand cDNA, transcriptor first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P

    Article Snippet: High pure RNA isolation kit and transcriptor first strand cDNA synthesis kit were obtained from Roche Diagnostics GmbH (Mannheim, Germany).

    Techniques: Isolation, Multiple Displacement Amplification, Real-time Polymerase Chain Reaction, SYBR Green Assay, Polymerase Chain Reaction, Expressing

    Evaluation of subtraction efficiency. A: Reduction of GAPDH cDNA following subtraction in the SH-Ctls sample. PCR was performed on SH-Ctls subtracted and SH unsubtracted samples. GAPDH PCR products (760 pb) were detectable 10 cycles earlier in the unsubtracted sample (18 cycles) than in the subtracted sample (28 cycles). B: Enrichment of LCN2 cDNA following subtraction in the SH-Ctls sample. PCR was performed on SH-Ctls and Ctls-SH subtracted samples as well as SH and Ctls unsubtracted samples. LCN2 PCR products (210 pb) were detected after 20 cycles for both SH unsubtracted and SH-Ctls subtracted samples, the difference in the intensity of the 2 bands indicate the enrichment compare to Ctls unsubtracted and Ctls-SH subtracted samples.

    Journal: PLoS ONE

    Article Title: Profiling of Differentially Expressed Genes Using Suppression Subtractive Hybridization in an Equine Model of Chronic Asthma

    doi: 10.1371/journal.pone.0029440

    Figure Lengend Snippet: Evaluation of subtraction efficiency. A: Reduction of GAPDH cDNA following subtraction in the SH-Ctls sample. PCR was performed on SH-Ctls subtracted and SH unsubtracted samples. GAPDH PCR products (760 pb) were detectable 10 cycles earlier in the unsubtracted sample (18 cycles) than in the subtracted sample (28 cycles). B: Enrichment of LCN2 cDNA following subtraction in the SH-Ctls sample. PCR was performed on SH-Ctls and Ctls-SH subtracted samples as well as SH and Ctls unsubtracted samples. LCN2 PCR products (210 pb) were detected after 20 cycles for both SH unsubtracted and SH-Ctls subtracted samples, the difference in the intensity of the 2 bands indicate the enrichment compare to Ctls unsubtracted and Ctls-SH subtracted samples.

    Article Snippet: First strand cDNA were generated using the SMART PCR cDNA Synthesis Kit (Clontech Laboratories, Inc., Mountain View, CA, USA) and the SuperScript III Reverse Transcriptase (Invitrogen, Carlsbad, CA, USA) for each individual horse at baseline and after challenge as described above.

    Techniques: Polymerase Chain Reaction

    Validation of alternative splicing events co-regulated by JMJD6 and U2AF65 through RT-PCR. ( A–J ) First-strand cDNA synthesis was done from RNA samples described in Figure 1B followed by standard PCR analysis (RT-PCR) using primer sets specifically targeting to the upstream and downstream exon relative to the cassette exon (alternatively spliced exon) to detect the expression of both short and long isoforms of representative genes as indicated. The length of the alternatively spliced exon as well as the expected length of the PCR products was shown as indicated. DNA marker (M) was included on the left. Percentage spliced in (PSI) was calculated as the ratio of the density of the long isoform versus that of the sum of the long and short isoforms. The position of the alternatively spliced exon in each gene was as follows: KIAA0515 (NM_013318, exon 30) (A); TCERG1 (BI091338, exon 3) (B); BAT2D1 (AV650960, exon 6) (C); HNRNPH3 (NM_012207, exon 3) (D); UTRN (NM_007124, exon 66) (E); TMEM126B (NM_018480, exon 2) (F); ZDHHC20 (uc001uod.1, exon 13) (G); BRD8 (NM_006696, exon 21) (H); PTBLP (NM_021190, exon 10) (I); NCAM1 (NM_181351, exon 9) (J). F: forward primer; R: reverse primer; bp: base pair.

    Journal: Nucleic Acids Research

    Article Title: JMJD6 and U2AF65 co-regulate alternative splicing in both JMJD6 enzymatic activity dependent and independent manner

    doi: 10.1093/nar/gkw1144

    Figure Lengend Snippet: Validation of alternative splicing events co-regulated by JMJD6 and U2AF65 through RT-PCR. ( A–J ) First-strand cDNA synthesis was done from RNA samples described in Figure 1B followed by standard PCR analysis (RT-PCR) using primer sets specifically targeting to the upstream and downstream exon relative to the cassette exon (alternatively spliced exon) to detect the expression of both short and long isoforms of representative genes as indicated. The length of the alternatively spliced exon as well as the expected length of the PCR products was shown as indicated. DNA marker (M) was included on the left. Percentage spliced in (PSI) was calculated as the ratio of the density of the long isoform versus that of the sum of the long and short isoforms. The position of the alternatively spliced exon in each gene was as follows: KIAA0515 (NM_013318, exon 30) (A); TCERG1 (BI091338, exon 3) (B); BAT2D1 (AV650960, exon 6) (C); HNRNPH3 (NM_012207, exon 3) (D); UTRN (NM_007124, exon 66) (E); TMEM126B (NM_018480, exon 2) (F); ZDHHC20 (uc001uod.1, exon 13) (G); BRD8 (NM_006696, exon 21) (H); PTBLP (NM_021190, exon 10) (I); NCAM1 (NM_181351, exon 9) (J). F: forward primer; R: reverse primer; bp: base pair.

    Article Snippet: First-strand cDNA synthesis from total RNA was carried out using iScript™ cDNA Synthesis Kit (Bio-Rad).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Expressing, Marker

    Adrenomedullin expression by breast cancer cell lines and effects of adrenomedullin on tumor and bone cells. (A) Adrenomedullin ( AM) RNA in human breast cancer cell lines that metastasize to bone. PCR detection of human AM mRNA in from left to right: normal tissues (kidney, prostate and breast), breast cancer cell lines (MCF-7, BT549, MDA-231, MCF-7, MDA-MB-436, ZR-75-1, T47D, MD-MB-361 and BT438) and three additional cancer cell lines that cause osteolytic bone metastases: MDA-MB-435S melanoma cells, PC-3 prostate cancer cells and A549 lung adenocarcinoma cells. cDNA (200 ng) was amplified for 30 cycles. The PCR product is 600 bp. It is unclear if kidney expresses only low amounts of AM mRNA or if the RNA was degraded. (B) Stable overexpression of human AM mRNA in MDA-MB-231 cells. Clones were tested for stable human AM mRNA expression using real-time PCR after 60 days of culture in the absence of antibiotic selection. G8-G31 are MDA-MB-231 control clones, and A15-A51 are MDA-MB-231 clones overexpressing AM. (C) Proliferation in vitro of breast cancer cells. Proliferation of two AM-overexpressing clones (A39, gray triangle; A51, black triangle) and two control clones (G8, gray square; G26, black square) were compared using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; tetrazolium blue dye) assay. Proliferation was compared after 1, 3 and 5 days of growth. OD, Optical density.

    Journal: Breast Cancer Research : BCR

    Article Title: Tumor-expressed adrenomedullin accelerates breast cancer bone metastasis

    doi: 10.1186/s13058-014-0458-y

    Figure Lengend Snippet: Adrenomedullin expression by breast cancer cell lines and effects of adrenomedullin on tumor and bone cells. (A) Adrenomedullin ( AM) RNA in human breast cancer cell lines that metastasize to bone. PCR detection of human AM mRNA in from left to right: normal tissues (kidney, prostate and breast), breast cancer cell lines (MCF-7, BT549, MDA-231, MCF-7, MDA-MB-436, ZR-75-1, T47D, MD-MB-361 and BT438) and three additional cancer cell lines that cause osteolytic bone metastases: MDA-MB-435S melanoma cells, PC-3 prostate cancer cells and A549 lung adenocarcinoma cells. cDNA (200 ng) was amplified for 30 cycles. The PCR product is 600 bp. It is unclear if kidney expresses only low amounts of AM mRNA or if the RNA was degraded. (B) Stable overexpression of human AM mRNA in MDA-MB-231 cells. Clones were tested for stable human AM mRNA expression using real-time PCR after 60 days of culture in the absence of antibiotic selection. G8-G31 are MDA-MB-231 control clones, and A15-A51 are MDA-MB-231 clones overexpressing AM. (C) Proliferation in vitro of breast cancer cells. Proliferation of two AM-overexpressing clones (A39, gray triangle; A51, black triangle) and two control clones (G8, gray square; G26, black square) were compared using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; tetrazolium blue dye) assay. Proliferation was compared after 1, 3 and 5 days of growth. OD, Optical density.

    Article Snippet: First-strand cDNA for PCR was made with an iScript cDNA synthesis kit (Bio-Rad Laboratories, Hercules, CA, USA).

    Techniques: Expressing, Polymerase Chain Reaction, Multiple Displacement Amplification, Amplification, Over Expression, Clone Assay, Real-time Polymerase Chain Reaction, Selection, In Vitro, MTT Assay