strand cdna synthesis Search Results


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  • 99
    Thermo Fisher first strand cdna synthesis
    Expression analysis of REM1 . A, An <t>RNA</t> blot containing 1.5 μg of poly(A) + RNA from different organs of Arabidopsis was hybridized with the <t>cDNA</t> REM1 and a probe corresponding to ribosomal protein L3 as a control for gel loading ( RBP4 ). B, Expression analysis of AtREM1 and AP2 by reverse transcriptase (RT)-PCR in different organs from adult plants. RT-PCR products were blotted onto membranes and hybridized with the corresponding probes. C, RT-PCR analysis of expression of REM1 in apices from young seedlings at different stages of development. C, Cotyledon and hypocotyl from young seedlings; IA, inflorescence apex; IF, immature flower, before anthesis; L, leaf; MF, flower at anthesis; R, root; S, stem; Sq, silique.
    First Strand Cdna Synthesis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 29586 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Roche strand complementary dna cdna synthesis kit
    Expression analysis of REM1 . A, An <t>RNA</t> blot containing 1.5 μg of poly(A) + RNA from different organs of Arabidopsis was hybridized with the <t>cDNA</t> REM1 and a probe corresponding to ribosomal protein L3 as a control for gel loading ( RBP4 ). B, Expression analysis of AtREM1 and AP2 by reverse transcriptase (RT)-PCR in different organs from adult plants. RT-PCR products were blotted onto membranes and hybridized with the corresponding probes. C, RT-PCR analysis of expression of REM1 in apices from young seedlings at different stages of development. C, Cotyledon and hypocotyl from young seedlings; IA, inflorescence apex; IF, immature flower, before anthesis; L, leaf; MF, flower at anthesis; R, root; S, stem; Sq, silique.
    Strand Complementary Dna Cdna Synthesis Kit, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GE Healthcare first strand cdna synthesis
    Expression analysis of REM1 . A, An <t>RNA</t> blot containing 1.5 μg of poly(A) + RNA from different organs of Arabidopsis was hybridized with the <t>cDNA</t> REM1 and a probe corresponding to ribosomal protein L3 as a control for gel loading ( RBP4 ). B, Expression analysis of AtREM1 and AP2 by reverse transcriptase (RT)-PCR in different organs from adult plants. RT-PCR products were blotted onto membranes and hybridized with the corresponding probes. C, RT-PCR analysis of expression of REM1 in apices from young seedlings at different stages of development. C, Cotyledon and hypocotyl from young seedlings; IA, inflorescence apex; IF, immature flower, before anthesis; L, leaf; MF, flower at anthesis; R, root; S, stem; Sq, silique.
    First Strand Cdna Synthesis, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 751 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Roche first strand complementary dna cdna synthesis kit
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    First Strand Complementary Dna Cdna Synthesis Kit, supplied by Roche, used in various techniques. Bioz Stars score: 88/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega strand complementary dna synthesis kit
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    Strand Complementary Dna Synthesis Kit, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Roche transcriptor first strand complementary dna cdna synthesis kit
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    Transcriptor First Strand Complementary Dna Cdna Synthesis Kit, supplied by Roche, used in various techniques. Bioz Stars score: 88/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    OriGene first strand complementary dna cdna synthesis system
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    First Strand Complementary Dna Cdna Synthesis System, supplied by OriGene, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher first strand complementary dna cdna synthesis
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    First Strand Complementary Dna Cdna Synthesis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 663 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    tiangen biotech co fastquant complementary dna cdna first strand synthesis kit
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    Fastquant Complementary Dna Cdna First Strand Synthesis Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    tiangen biotech co strand complementary dna cdna synthesis
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    Strand Complementary Dna Cdna Synthesis, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher double stranded complementary dna cdna synthesis kit
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    Double Stranded Complementary Dna Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Quanta Biosciences strand complementary dna cdna synthesis
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    Strand Complementary Dna Cdna Synthesis, supplied by Quanta Biosciences, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher first strand complementary dna cdna synthesis kit
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    First Strand Complementary Dna Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 89 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    TaKaRa complementary dna cdna synthesis primescripttm 1st strand cdna synthesis kit
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    Complementary Dna Cdna Synthesis Primescripttm 1st Strand Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad strand complementary dna cdna synthesis
    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by <t>DNA</t> sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using <t>cDNA</t> sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.
    Strand Complementary Dna Cdna Synthesis, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher cdna
    Decreased Egr-1 expression in kidneys from Fli-1 +/− NZM2410 mice compared to wild-type controls. Total <t>RNA</t> was prepared from kidneys at the age of 18 weeks ( n = 4 in each group). Total RNA was converted to <t>cDNA</t> with the SuperScript First-Strand
    Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 160058 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    GE Healthcare strand complementary dna cdna synthesis
    Decreased Egr-1 expression in kidneys from Fli-1 +/− NZM2410 mice compared to wild-type controls. Total <t>RNA</t> was prepared from kidneys at the age of 18 weeks ( n = 4 in each group). Total RNA was converted to <t>cDNA</t> with the SuperScript First-Strand
    Strand Complementary Dna Cdna Synthesis, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    GE Healthcare first strand cdna synthesis kit
    Translational induction of DAP5/p97 during ER stress is caspase-independent. ( A ) HEK293T cells were pre-treated with 100 μM Z-VAD-FMK or DMSO for 6 h and were then incubated with 8.5 μM tunicamycin or DMSO in the presence or absence of 100 μM Z-VAD-FMK for an additional 24 h. Protein extracts were harvested, separated by 10% SDS–PAGE, transferred to PVDF membrane and the levels of BiP, p97, GAPDH and cleaved PARP were determined by western blot analysis. ( B ) IRES activity of the DAP5/p97 and HIAP2 5′ UTRs was determined in DAP5/p97-overexpressing or control plasmid transfected cells using the bicistronic reporter plasmids described in Figure 3 A. Average ± SD of three independent experiments performed in triplicate. Samples from the control plasmid transfected cells were set as 1. ( C ) Both DAP5/p97 and HIAP2 mRNA associate with full-length DAP5/p97. HEK293T cells were transfected with pCI, FLAG- DAP5/p97 or FLAG- DAP5/p86 and mRNA:protein complexes were co-precipitated using anti-FLAG coated agarose beads as described in Materials and Methods section. <t>cDNA</t> was produced from precipitated mRNA by reverse transcription (RT) using an oligo d(T) 18 primer, which was subsequently amplified by <t>PCR</t> using gene-specific oligonucleotide primers for DAP5/p97, HIAP2 and GAPDH. The resulting products were separated on a 1.5% agarose gel and visualized by ethidium bromide staining.
    First Strand Cdna Synthesis Kit, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 94/100, based on 5249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    TaKaRa first strand complementary dna synthesis kit
    Translational induction of DAP5/p97 during ER stress is caspase-independent. ( A ) HEK293T cells were pre-treated with 100 μM Z-VAD-FMK or DMSO for 6 h and were then incubated with 8.5 μM tunicamycin or DMSO in the presence or absence of 100 μM Z-VAD-FMK for an additional 24 h. Protein extracts were harvested, separated by 10% SDS–PAGE, transferred to PVDF membrane and the levels of BiP, p97, GAPDH and cleaved PARP were determined by western blot analysis. ( B ) IRES activity of the DAP5/p97 and HIAP2 5′ UTRs was determined in DAP5/p97-overexpressing or control plasmid transfected cells using the bicistronic reporter plasmids described in Figure 3 A. Average ± SD of three independent experiments performed in triplicate. Samples from the control plasmid transfected cells were set as 1. ( C ) Both DAP5/p97 and HIAP2 mRNA associate with full-length DAP5/p97. HEK293T cells were transfected with pCI, FLAG- DAP5/p97 or FLAG- DAP5/p86 and mRNA:protein complexes were co-precipitated using anti-FLAG coated agarose beads as described in Materials and Methods section. <t>cDNA</t> was produced from precipitated mRNA by reverse transcription (RT) using an oligo d(T) 18 primer, which was subsequently amplified by <t>PCR</t> using gene-specific oligonucleotide primers for DAP5/p97, HIAP2 and GAPDH. The resulting products were separated on a 1.5% agarose gel and visualized by ethidium bromide staining.
    First Strand Complementary Dna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa first strand complementary dna cdna synthesis
    Translational induction of DAP5/p97 during ER stress is caspase-independent. ( A ) HEK293T cells were pre-treated with 100 μM Z-VAD-FMK or DMSO for 6 h and were then incubated with 8.5 μM tunicamycin or DMSO in the presence or absence of 100 μM Z-VAD-FMK for an additional 24 h. Protein extracts were harvested, separated by 10% SDS–PAGE, transferred to PVDF membrane and the levels of BiP, p97, GAPDH and cleaved PARP were determined by western blot analysis. ( B ) IRES activity of the DAP5/p97 and HIAP2 5′ UTRs was determined in DAP5/p97-overexpressing or control plasmid transfected cells using the bicistronic reporter plasmids described in Figure 3 A. Average ± SD of three independent experiments performed in triplicate. Samples from the control plasmid transfected cells were set as 1. ( C ) Both DAP5/p97 and HIAP2 mRNA associate with full-length DAP5/p97. HEK293T cells were transfected with pCI, FLAG- DAP5/p97 or FLAG- DAP5/p86 and mRNA:protein complexes were co-precipitated using anti-FLAG coated agarose beads as described in Materials and Methods section. <t>cDNA</t> was produced from precipitated mRNA by reverse transcription (RT) using an oligo d(T) 18 primer, which was subsequently amplified by <t>PCR</t> using gene-specific oligonucleotide primers for DAP5/p97, HIAP2 and GAPDH. The resulting products were separated on a 1.5% agarose gel and visualized by ethidium bromide staining.
    First Strand Complementary Dna Cdna Synthesis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Expression analysis of REM1 . A, An RNA blot containing 1.5 μg of poly(A) + RNA from different organs of Arabidopsis was hybridized with the cDNA REM1 and a probe corresponding to ribosomal protein L3 as a control for gel loading ( RBP4 ). B, Expression analysis of AtREM1 and AP2 by reverse transcriptase (RT)-PCR in different organs from adult plants. RT-PCR products were blotted onto membranes and hybridized with the corresponding probes. C, RT-PCR analysis of expression of REM1 in apices from young seedlings at different stages of development. C, Cotyledon and hypocotyl from young seedlings; IA, inflorescence apex; IF, immature flower, before anthesis; L, leaf; MF, flower at anthesis; R, root; S, stem; Sq, silique.

    Journal: Plant Physiology

    Article Title: AtREM1, a Member of a New Family of B3 Domain-Containing Genes, Is Preferentially Expressed in Reproductive Meristems 1

    doi: 10.1104/pp.010323

    Figure Lengend Snippet: Expression analysis of REM1 . A, An RNA blot containing 1.5 μg of poly(A) + RNA from different organs of Arabidopsis was hybridized with the cDNA REM1 and a probe corresponding to ribosomal protein L3 as a control for gel loading ( RBP4 ). B, Expression analysis of AtREM1 and AP2 by reverse transcriptase (RT)-PCR in different organs from adult plants. RT-PCR products were blotted onto membranes and hybridized with the corresponding probes. C, RT-PCR analysis of expression of REM1 in apices from young seedlings at different stages of development. C, Cotyledon and hypocotyl from young seedlings; IA, inflorescence apex; IF, immature flower, before anthesis; L, leaf; MF, flower at anthesis; R, root; S, stem; Sq, silique.

    Article Snippet: For RT-PCR expression studies in rem1 homozygous lines, 5 μg of total RNA from inflorescence apices was treated with DNase and was employed in first-strand cDNA synthesis using SuperScript II (Gibco-BRL, Grand Island, NY) and an oligo T17 as primer. cDNAs were 250- and 500-fold diluted in subsequent PCR reactions.

    Techniques: Expressing, Northern blot, Reverse Transcription Polymerase Chain Reaction, IA

    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

    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by DNA sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using cDNA sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.

    Journal: Medicine

    Article Title: Clinical and Molecular Characterization of NF1 Patients

    doi: 10.1097/MD.0000000000003043

    Figure Lengend Snippet: Flow chart for comprehensive NF1 mutation detection. Point mutations identified by DNA sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using cDNA sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.

    Article Snippet: To prevent illegitimate splicing, blood samples were processed after venipuncture with a maximum delay of 4 h and samples were not stored at 4°C., Reverse transcription was performed using 500 ng of total RNA isolated and random hexamers with a First-Strand complementary DNA (cDNA) Synthesis Kit for RT-PCR (AMV) (Roche Applied Science, Indianapolis, IN).

    Techniques: Flow Cytometry, Mutagenesis, DNA Sequencing, Sequencing, Multiplex Assay, Ligation, Amplification, Multiplex Ligation-dependent Probe Amplification

    Effects of different doses of the PPAR δ agonist, selective inhibitor of MEK/ERK1/2, and EGF receptor-selective tyrosine kinase inhibitor on mRNA expression of Δ6-desaturase (Δ6D) in PANC-1 human pancreatic tumor cells. Cells were treated with GW0742, PD98059, or AG1478 as indicated. One microgram of extracted RNA was reverse transcribed, and real-time RT-PCR to amplify Δ6 desaturase and GAPDH cDNA fragments was performed as described. Relative expression with duplicate samples is given and was calculated by normalization to GAPDH and represented as mean ± SE from 3 independent experiments. Data are means ± SE, n = 3. **: P

    Journal: The Scientific World Journal

    Article Title: Transcriptional Regulation of ?6-Desaturase by Peroxisome Proliferative-Activated Receptor δ Agonist in Human Pancreatic Cancer Cells: Role of MEK/ERK1/2 Pathway

    doi: 10.1155/2013/607524

    Figure Lengend Snippet: Effects of different doses of the PPAR δ agonist, selective inhibitor of MEK/ERK1/2, and EGF receptor-selective tyrosine kinase inhibitor on mRNA expression of Δ6-desaturase (Δ6D) in PANC-1 human pancreatic tumor cells. Cells were treated with GW0742, PD98059, or AG1478 as indicated. One microgram of extracted RNA was reverse transcribed, and real-time RT-PCR to amplify Δ6 desaturase and GAPDH cDNA fragments was performed as described. Relative expression with duplicate samples is given and was calculated by normalization to GAPDH and represented as mean ± SE from 3 independent experiments. Data are means ± SE, n = 3. **: P

    Article Snippet: For cDNA synthesis, RNA (1 μ g) was reverse transcribed with a first-strand cDNA synthesis kit for reverse-transcription polymerase chain reaction (RT-PCR; Roche, Hertfordshire, UK).

    Techniques: Expressing, Quantitative RT-PCR

    Effects of the PPAR δ agonist, selective inhibitor of MEK/ERK1/2, and EGF receptor-selective tyrosine kinase inhibitor on mRNA expression of Δ6-desaturase (Δ6D) in PANC-1 human pancreatic tumor cells. Cells were treated with GW0742, PD98059, or AG1478 as indicated. One microgram of extracted RNA was reverse transcribed, and real-time RT-PCR to amplify Δ6 desaturase and GAPDH cDNA fragments was performed as described. Relative expression with duplicate samples is given and was calculated by normalization to GAPDH and represented as mean ± SE from 3 independent experiments. * and **: P

    Journal: The Scientific World Journal

    Article Title: Transcriptional Regulation of ?6-Desaturase by Peroxisome Proliferative-Activated Receptor δ Agonist in Human Pancreatic Cancer Cells: Role of MEK/ERK1/2 Pathway

    doi: 10.1155/2013/607524

    Figure Lengend Snippet: Effects of the PPAR δ agonist, selective inhibitor of MEK/ERK1/2, and EGF receptor-selective tyrosine kinase inhibitor on mRNA expression of Δ6-desaturase (Δ6D) in PANC-1 human pancreatic tumor cells. Cells were treated with GW0742, PD98059, or AG1478 as indicated. One microgram of extracted RNA was reverse transcribed, and real-time RT-PCR to amplify Δ6 desaturase and GAPDH cDNA fragments was performed as described. Relative expression with duplicate samples is given and was calculated by normalization to GAPDH and represented as mean ± SE from 3 independent experiments. * and **: P

    Article Snippet: For cDNA synthesis, RNA (1 μ g) was reverse transcribed with a first-strand cDNA synthesis kit for reverse-transcription polymerase chain reaction (RT-PCR; Roche, Hertfordshire, UK).

    Techniques: Expressing, Quantitative RT-PCR

    Decreased Egr-1 expression in kidneys from Fli-1 +/− NZM2410 mice compared to wild-type controls. Total RNA was prepared from kidneys at the age of 18 weeks ( n = 4 in each group). Total RNA was converted to cDNA with the SuperScript First-Strand

    Journal: Clinical and Experimental Immunology

    Article Title: Impact of Fli-1 transcription factor on autoantibody and lupus nephritis in NZM2410 mice

    doi: 10.1111/j.1365-2249.2010.04245.x

    Figure Lengend Snippet: Decreased Egr-1 expression in kidneys from Fli-1 +/− NZM2410 mice compared to wild-type controls. Total RNA was prepared from kidneys at the age of 18 weeks ( n = 4 in each group). Total RNA was converted to cDNA with the SuperScript First-Strand

    Article Snippet: Two µg of RNA was used to synthesize cDNA (SuperScript First-Strand Synthesis System; Invitrogen).

    Techniques: Expressing, Mouse Assay

    Translational induction of DAP5/p97 during ER stress is caspase-independent. ( A ) HEK293T cells were pre-treated with 100 μM Z-VAD-FMK or DMSO for 6 h and were then incubated with 8.5 μM tunicamycin or DMSO in the presence or absence of 100 μM Z-VAD-FMK for an additional 24 h. Protein extracts were harvested, separated by 10% SDS–PAGE, transferred to PVDF membrane and the levels of BiP, p97, GAPDH and cleaved PARP were determined by western blot analysis. ( B ) IRES activity of the DAP5/p97 and HIAP2 5′ UTRs was determined in DAP5/p97-overexpressing or control plasmid transfected cells using the bicistronic reporter plasmids described in Figure 3 A. Average ± SD of three independent experiments performed in triplicate. Samples from the control plasmid transfected cells were set as 1. ( C ) Both DAP5/p97 and HIAP2 mRNA associate with full-length DAP5/p97. HEK293T cells were transfected with pCI, FLAG- DAP5/p97 or FLAG- DAP5/p86 and mRNA:protein complexes were co-precipitated using anti-FLAG coated agarose beads as described in Materials and Methods section. cDNA was produced from precipitated mRNA by reverse transcription (RT) using an oligo d(T) 18 primer, which was subsequently amplified by PCR using gene-specific oligonucleotide primers for DAP5/p97, HIAP2 and GAPDH. The resulting products were separated on a 1.5% agarose gel and visualized by ethidium bromide staining.

    Journal: Nucleic Acids Research

    Article Title: The eIF4G homolog DAP5/p97 supports the translation of select mRNAs during endoplasmic reticulum stress

    doi: 10.1093/nar/gkm1007

    Figure Lengend Snippet: Translational induction of DAP5/p97 during ER stress is caspase-independent. ( A ) HEK293T cells were pre-treated with 100 μM Z-VAD-FMK or DMSO for 6 h and were then incubated with 8.5 μM tunicamycin or DMSO in the presence or absence of 100 μM Z-VAD-FMK for an additional 24 h. Protein extracts were harvested, separated by 10% SDS–PAGE, transferred to PVDF membrane and the levels of BiP, p97, GAPDH and cleaved PARP were determined by western blot analysis. ( B ) IRES activity of the DAP5/p97 and HIAP2 5′ UTRs was determined in DAP5/p97-overexpressing or control plasmid transfected cells using the bicistronic reporter plasmids described in Figure 3 A. Average ± SD of three independent experiments performed in triplicate. Samples from the control plasmid transfected cells were set as 1. ( C ) Both DAP5/p97 and HIAP2 mRNA associate with full-length DAP5/p97. HEK293T cells were transfected with pCI, FLAG- DAP5/p97 or FLAG- DAP5/p86 and mRNA:protein complexes were co-precipitated using anti-FLAG coated agarose beads as described in Materials and Methods section. cDNA was produced from precipitated mRNA by reverse transcription (RT) using an oligo d(T) 18 primer, which was subsequently amplified by PCR using gene-specific oligonucleotide primers for DAP5/p97, HIAP2 and GAPDH. The resulting products were separated on a 1.5% agarose gel and visualized by ethidium bromide staining.

    Article Snippet: For quantitative RT-PCR, reverse transcription was carried out using the First-Strand cDNA Synthesis kit (Amersham Biosciences, Piscataway, NJ, USA) with oligo d(T)18 primers.

    Techniques: Incubation, SDS Page, Western Blot, Activity Assay, Plasmid Preparation, Transfection, Produced, Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining