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    Structured Review

    HyTest dntp
    Dntp, supplied by HyTest, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dntp/product/HyTest
    Average 90 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    dntp - by Bioz Stars, 2020-04
    90/100 stars

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    Related Articles

    Diagnostic Assay:

    Article Title: Screening of the DNA mismatch repair genes MLH1, MSH2 and MSH6 in a Greek cohort of Lynch syndrome suspected families
    Article Snippet: .. PCR using diagnostic primers 100 ng of genomic DNA was amplified in a 20 μl reaction volume using 1.5 mM Mg+2 , 100 μM of each dNTP, 1.25 U of Taq polymerase (HyTest Ltd. Intelligate, Finland). .. Two primer sets were used (F3/R1 & For/Rev ×15 BRAF) in order to achieve the simultaneous amplification of the locus involving the exon 6 deletion and an independent locus used as an internal control [ , ].

    Real-time Polymerase Chain Reaction:

    Article Title: Microarray Analysis of mRNA and MicroRNA Expression Profile Reveals the Role of β-Sitosterol-D-glucoside in the Proliferation of Neural Stem Cell
    Article Snippet: Paragraph title: 2.5.2. Real-Time PCR Validation for miRNA Expression Profiling ... Total RNA was reverse-transcribed to cDNA using AMV reverse transcriptase (Epicentre), RNase (Epicentre), dNTP (HyTest Ltd), RT buffer, and RT primers (Invitrogen).

    Article Title: Effects of Folic Acid on Secretases Involved in Aβ Deposition in APP/PS1 Mice
    Article Snippet: Paragraph title: 2.6. Real-Time PCR for mRNA and miRNA Expression ... For miRNA determination, total RNA was reverse-transcribed to cDNA using RNase Inhibitor (Epicentre, Madison, WI, USA), dNTP (HyTest Ltd., Turku, Finland), RT buffer, and RT primers (Invitrogen).

    Article Title: Differential miRNA expression profiles in human keratinocytes in response to protein kinase C inhibitor
    Article Snippet: RT was performed in a 20 µl reaction containing 200 ng total RNA, 0.3 µl 1 µM RT primer, 2 µl 2.5 mM dNTP (HyTest Ltd, Turku, Finland), 2 µl 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µl 50 U/µl RT enzyme (Epicentre; Illumina, Inc.), 0.3 µl 40 U/µl RNase inhibitor (Epicentre; Illumina, Inc.), 20 µl nuclease free water and 0.2 µl MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.). .. The stem-loop RT reaction was performed at 16°C for 30 min, followed by 42°C for 30 min and 85°C for 5 min. A total of 2 µl RT reaction was then used with 1 µl specific primers for each of the hsa-miR-1-3p and hsa-miR-31-5p in triplicate wells for PCR on an Applied Biosystems ViiA 7 Real-time PCR system (Applied Biosystems, Thermo Fisher Scientific, Inc.).

    Amplification:

    Article Title: Effects of Folic Acid on Secretases Involved in Aβ Deposition in APP/PS1 Mice
    Article Snippet: The reaction mixtures were incubated at 95 °C for 5 min, which was followed by 45 amplification cycles (denaturation, 95 °C for 10 s; annealing, 56 °C for ADAM9 and ADAM10, 59 °C for BACE1 and 63 °C for PS1 for 10 s; extension, 72 °C for 10 s). .. For miRNA determination, total RNA was reverse-transcribed to cDNA using RNase Inhibitor (Epicentre, Madison, WI, USA), dNTP (HyTest Ltd., Turku, Finland), RT buffer, and RT primers (Invitrogen).

    Article Title: Screening of the DNA mismatch repair genes MLH1, MSH2 and MSH6 in a Greek cohort of Lynch syndrome suspected families
    Article Snippet: .. PCR using diagnostic primers 100 ng of genomic DNA was amplified in a 20 μl reaction volume using 1.5 mM Mg+2 , 100 μM of each dNTP, 1.25 U of Taq polymerase (HyTest Ltd. Intelligate, Finland). .. Two primer sets were used (F3/R1 & For/Rev ×15 BRAF) in order to achieve the simultaneous amplification of the locus involving the exon 6 deletion and an independent locus used as an internal control [ , ].

    Article Title: Polarisation of Major Histocompatibility Complex II Host Genotype with Pathogenesis of European Brown Hare Syndrome Virus
    Article Snippet: Exon 2 locus of hare MHC DQA gene was amplified using the primers DQA-Fw:5′-GTTGGTGCCTATGGCATAAA-3′, DQA-Rv: 5′-AGCAGTAGAGTTGGAGCGTTT-3′ with slight modifications. .. PCR reactions (30 µL) contained 200–500 ng of genomic DNA, 10× Taq buffer, 2 mM MgCl2 , 0.2 mM of each dNTP, 25 pmoles of each primer and 1 U Taq polymerase (HyTest).

    Synthesized:

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: The sequences of PCR primers are listed in , which was designed by Primer 5.0 (Premier Biosoft International, Palo Alto, CA, USA) Firstly, cDNA was synthesized by using a Gene Amp PCR system 9700 (Applied Biosystems; Thermo Fisher Scientific, Inc.). .. RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Article Title: MicroRNA profile comparison of the corneal endothelia of young and old mice: implications for senescence of the corneal endothelium
    Article Snippet: All specific primers were designed and synthesized by Guangzhou RiboBio (RiboBio, Guangzhou, China) using the qRT-PCR Primer Sets (catalog no. MQP-0101). .. Briefly, in a 20 μl reaction volume, 1 μM of each specific reverse transcription primer (0.3 μl), 2 μl dNTP (2.5mM; HyTest Ltd., Turku, Finland), 2 μl 10X RT buffer (Promega Corporation, Madison, WI), 20 U MMLV Reverse Transcriptase (10 U/μl; Epicenter Biotechnologies), 0.3 μL RNase inhibitor (40 U/μl), and 1 μg of total RNA was added. qRT-PCR was performed using the SYBR Green protocol on an ABI 7500 system (Applied Biosystems, Foster City, CA), and a data analysis was performed with the SDS system software (7500 System; Applied Biosystems).

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: The sequences of PCR primers are listed in , which was designed by Primer 5.0 (Premier Biosoft International, Palo Alto, CA, USA) Firstly, cDNA was synthesized by using a Gene Amp PCR system 9700 (Applied Biosystems; Thermo Fisher Scientific, Inc.). .. RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/ µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/ µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Article Title: Differential miRNA expression profiles in human keratinocytes in response to protein kinase C inhibitor
    Article Snippet: Firstly, cDNA was synthesized using a Gene Amp PCR system 9700 (Applied Biosystems, Thermo Fisher Scientific, Inc.). .. RT was performed in a 20 µl reaction containing 200 ng total RNA, 0.3 µl 1 µM RT primer, 2 µl 2.5 mM dNTP (HyTest Ltd, Turku, Finland), 2 µl 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µl 50 U/µl RT enzyme (Epicentre; Illumina, Inc.), 0.3 µl 40 U/µl RNase inhibitor (Epicentre; Illumina, Inc.), 20 µl nuclease free water and 0.2 µl MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Sequencing:

    Article Title: Effects of Folic Acid on Secretases Involved in Aβ Deposition in APP/PS1 Mice
    Article Snippet: The assay was performed using the Roche LightCycler 480 sequence detector (Roche). .. For miRNA determination, total RNA was reverse-transcribed to cDNA using RNase Inhibitor (Epicentre, Madison, WI, USA), dNTP (HyTest Ltd., Turku, Finland), RT buffer, and RT primers (Invitrogen).

    Size-exclusion Chromatography:

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.). .. The stem-loop RT reaction was performed at 16°C for 30 min, followed by 42°C for 30 min and 85°C for 5 min. Then, RT-qPCR was performed using the SYBR Green I RT-PCR kit (Life Sciences; Thermo Fisher Scientific, Inc.) and GAPDH was used as an internal control, with the following reaction profile: Predenaturation at 95°C for 10 sec, followed by 40 cycles of 95°C for 10 sec and 60°C for 1 min.

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/ µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/ µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.). .. The stem-loop RT reaction was performed at 16°C for 30 min, followed by 42°C for 30 min and 85°C for 5 min. Then, RT-qPCR was performed using the SYBR Green I RT-PCR kit (Life Sciences; Thermo Fisher Scientific, Inc.) and GAPDH was used as an internal control, with the following reaction profile: Predenaturation at 95°C for 10 sec, followed by 40 cycles of 95°C for 10 sec and 60°C for 1 min.

    Article Title: Polarisation of Major Histocompatibility Complex II Host Genotype with Pathogenesis of European Brown Hare Syndrome Virus
    Article Snippet: PCR reactions (30 µL) contained 200–500 ng of genomic DNA, 10× Taq buffer, 2 mM MgCl2 , 0.2 mM of each dNTP, 25 pmoles of each primer and 1 U Taq polymerase (HyTest). .. The cycling conditions consisted of an initial denaturation at 95°C for 4 min followed by 35 rounds of denaturation at 95°C for 30 sec, annealing at 58°C for 40 sec and extension at 72°C for 40 sec, with a final extension at 72°C for 10 min using an Eppendorf thermal cycler.

    Article Title: Differential miRNA expression profiles in human keratinocytes in response to protein kinase C inhibitor
    Article Snippet: RT was performed in a 20 µl reaction containing 200 ng total RNA, 0.3 µl 1 µM RT primer, 2 µl 2.5 mM dNTP (HyTest Ltd, Turku, Finland), 2 µl 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µl 50 U/µl RT enzyme (Epicentre; Illumina, Inc.), 0.3 µl 40 U/µl RNase inhibitor (Epicentre; Illumina, Inc.), 20 µl nuclease free water and 0.2 µl MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.). .. The thermal cycling parameters were as follows: An initial predenaturation step at 95°C for 10 min, followed by 40 cycles of denaturation at 95°C for 10 sec, annealing at 60°C for 60 sec, followed by 95°C for 10 sec, 60°C for 60 sec and 95°C for 15 sec.

    Quantitative RT-PCR:

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: Paragraph title: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) ... RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Article Title: MicroRNA profile comparison of the corneal endothelia of young and old mice: implications for senescence of the corneal endothelium
    Article Snippet: .. Briefly, in a 20 μl reaction volume, 1 μM of each specific reverse transcription primer (0.3 μl), 2 μl dNTP (2.5mM; HyTest Ltd., Turku, Finland), 2 μl 10X RT buffer (Promega Corporation, Madison, WI), 20 U MMLV Reverse Transcriptase (10 U/μl; Epicenter Biotechnologies), 0.3 μL RNase inhibitor (40 U/μl), and 1 μg of total RNA was added. qRT-PCR was performed using the SYBR Green protocol on an ABI 7500 system (Applied Biosystems, Foster City, CA), and a data analysis was performed with the SDS system software (7500 System; Applied Biosystems). .. The levels of an endogenous control, U6 (RiboBio, Guangzhou, China; catalog no. MQP-0201), were used to normalize the expression levels of each miRNA.

    Article Title: Differential miRNA expression profiles in human keratinocytes in response to protein kinase C inhibitor
    Article Snippet: Paragraph title: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and miRNA target prediction ... RT was performed in a 20 µl reaction containing 200 ng total RNA, 0.3 µl 1 µM RT primer, 2 µl 2.5 mM dNTP (HyTest Ltd, Turku, Finland), 2 µl 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µl 50 U/µl RT enzyme (Epicentre; Illumina, Inc.), 0.3 µl 40 U/µl RNase inhibitor (Epicentre; Illumina, Inc.), 20 µl nuclease free water and 0.2 µl MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Purification:

    Article Title: Polarisation of Major Histocompatibility Complex II Host Genotype with Pathogenesis of European Brown Hare Syndrome Virus
    Article Snippet: PCR reactions (30 µL) contained 200–500 ng of genomic DNA, 10× Taq buffer, 2 mM MgCl2 , 0.2 mM of each dNTP, 25 pmoles of each primer and 1 U Taq polymerase (HyTest). .. PCR products showing the same SSCP pattern were grouped and representative samples from each profile were purified with Qiagen purification kit.

    SYBR Green Assay:

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.). .. The stem-loop RT reaction was performed at 16°C for 30 min, followed by 42°C for 30 min and 85°C for 5 min. Then, RT-qPCR was performed using the SYBR Green I RT-PCR kit (Life Sciences; Thermo Fisher Scientific, Inc.) and GAPDH was used as an internal control, with the following reaction profile: Predenaturation at 95°C for 10 sec, followed by 40 cycles of 95°C for 10 sec and 60°C for 1 min.

    Article Title: MicroRNA profile comparison of the corneal endothelia of young and old mice: implications for senescence of the corneal endothelium
    Article Snippet: .. Briefly, in a 20 μl reaction volume, 1 μM of each specific reverse transcription primer (0.3 μl), 2 μl dNTP (2.5mM; HyTest Ltd., Turku, Finland), 2 μl 10X RT buffer (Promega Corporation, Madison, WI), 20 U MMLV Reverse Transcriptase (10 U/μl; Epicenter Biotechnologies), 0.3 μL RNase inhibitor (40 U/μl), and 1 μg of total RNA was added. qRT-PCR was performed using the SYBR Green protocol on an ABI 7500 system (Applied Biosystems, Foster City, CA), and a data analysis was performed with the SDS system software (7500 System; Applied Biosystems). .. The levels of an endogenous control, U6 (RiboBio, Guangzhou, China; catalog no. MQP-0201), were used to normalize the expression levels of each miRNA.

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/ µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/ µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.). .. The stem-loop RT reaction was performed at 16°C for 30 min, followed by 42°C for 30 min and 85°C for 5 min. Then, RT-qPCR was performed using the SYBR Green I RT-PCR kit (Life Sciences; Thermo Fisher Scientific, Inc.) and GAPDH was used as an internal control, with the following reaction profile: Predenaturation at 95°C for 10 sec, followed by 40 cycles of 95°C for 10 sec and 60°C for 1 min.

    Polymerase Chain Reaction:

    Article Title: Effects of Folic Acid on Secretases Involved in Aβ Deposition in APP/PS1 Mice
    Article Snippet: The 20 μL PCR mixture included 10 μL of PCR Master mix, 5 μL of cDNA, 1 μL of forward primer, 1 μL of reverse primer, and 3 μL of water. .. For miRNA determination, total RNA was reverse-transcribed to cDNA using RNase Inhibitor (Epicentre, Madison, WI, USA), dNTP (HyTest Ltd., Turku, Finland), RT buffer, and RT primers (Invitrogen).

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: Paragraph title: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) ... RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Article Title: Screening of the DNA mismatch repair genes MLH1, MSH2 and MSH6 in a Greek cohort of Lynch syndrome suspected families
    Article Snippet: .. PCR using diagnostic primers 100 ng of genomic DNA was amplified in a 20 μl reaction volume using 1.5 mM Mg+2 , 100 μM of each dNTP, 1.25 U of Taq polymerase (HyTest Ltd. Intelligate, Finland). .. Two primer sets were used (F3/R1 & For/Rev ×15 BRAF) in order to achieve the simultaneous amplification of the locus involving the exon 6 deletion and an independent locus used as an internal control [ , ].

    Article Title: Polarisation of Major Histocompatibility Complex II Host Genotype with Pathogenesis of European Brown Hare Syndrome Virus
    Article Snippet: .. PCR reactions (30 µL) contained 200–500 ng of genomic DNA, 10× Taq buffer, 2 mM MgCl2 , 0.2 mM of each dNTP, 25 pmoles of each primer and 1 U Taq polymerase (HyTest). .. The cycling conditions consisted of an initial denaturation at 95°C for 4 min followed by 35 rounds of denaturation at 95°C for 30 sec, annealing at 58°C for 40 sec and extension at 72°C for 40 sec, with a final extension at 72°C for 10 min using an Eppendorf thermal cycler.

    Article Title: Differential miRNA expression profiles in human keratinocytes in response to protein kinase C inhibitor
    Article Snippet: Paragraph title: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and miRNA target prediction ... RT was performed in a 20 µl reaction containing 200 ng total RNA, 0.3 µl 1 µM RT primer, 2 µl 2.5 mM dNTP (HyTest Ltd, Turku, Finland), 2 µl 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µl 50 U/µl RT enzyme (Epicentre; Illumina, Inc.), 0.3 µl 40 U/µl RNase inhibitor (Epicentre; Illumina, Inc.), 20 µl nuclease free water and 0.2 µl MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Incubation:

    Article Title: Microarray Analysis of mRNA and MicroRNA Expression Profile Reveals the Role of β-Sitosterol-D-glucoside in the Proliferation of Neural Stem Cell
    Article Snippet: Total RNA was reverse-transcribed to cDNA using AMV reverse transcriptase (Epicentre), RNase (Epicentre), dNTP (HyTest Ltd), RT buffer, and RT primers (Invitrogen). .. The mixture was incubated at 16°C for 30 min, 42°C for 40 min, and 85°C for 5 min to generate a library of miRNA cDNAs.

    Article Title: Effects of Folic Acid on Secretases Involved in Aβ Deposition in APP/PS1 Mice
    Article Snippet: The reaction mixtures were incubated at 95 °C for 5 min, which was followed by 45 amplification cycles (denaturation, 95 °C for 10 s; annealing, 56 °C for ADAM9 and ADAM10, 59 °C for BACE1 and 63 °C for PS1 for 10 s; extension, 72 °C for 10 s). .. For miRNA determination, total RNA was reverse-transcribed to cDNA using RNase Inhibitor (Epicentre, Madison, WI, USA), dNTP (HyTest Ltd., Turku, Finland), RT buffer, and RT primers (Invitrogen).

    Expressing:

    Article Title: Microarray Analysis of mRNA and MicroRNA Expression Profile Reveals the Role of β-Sitosterol-D-glucoside in the Proliferation of Neural Stem Cell
    Article Snippet: Paragraph title: 2.5.2. Real-Time PCR Validation for miRNA Expression Profiling ... Total RNA was reverse-transcribed to cDNA using AMV reverse transcriptase (Epicentre), RNase (Epicentre), dNTP (HyTest Ltd), RT buffer, and RT primers (Invitrogen).

    Article Title: Effects of Folic Acid on Secretases Involved in Aβ Deposition in APP/PS1 Mice
    Article Snippet: Paragraph title: 2.6. Real-Time PCR for mRNA and miRNA Expression ... For miRNA determination, total RNA was reverse-transcribed to cDNA using RNase Inhibitor (Epicentre, Madison, WI, USA), dNTP (HyTest Ltd., Turku, Finland), RT buffer, and RT primers (Invitrogen).

    Article Title: MicroRNA profile comparison of the corneal endothelia of young and old mice: implications for senescence of the corneal endothelium
    Article Snippet: Briefly, in a 20 μl reaction volume, 1 μM of each specific reverse transcription primer (0.3 μl), 2 μl dNTP (2.5mM; HyTest Ltd., Turku, Finland), 2 μl 10X RT buffer (Promega Corporation, Madison, WI), 20 U MMLV Reverse Transcriptase (10 U/μl; Epicenter Biotechnologies), 0.3 μL RNase inhibitor (40 U/μl), and 1 μg of total RNA was added. qRT-PCR was performed using the SYBR Green protocol on an ABI 7500 system (Applied Biosystems, Foster City, CA), and a data analysis was performed with the SDS system software (7500 System; Applied Biosystems). .. The levels of an endogenous control, U6 (RiboBio, Guangzhou, China; catalog no. MQP-0201), were used to normalize the expression levels of each miRNA.

    Article Title: Differential miRNA expression profiles in human keratinocytes in response to protein kinase C inhibitor
    Article Snippet: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and miRNA target prediction According to the microarray results, the expression levels of hsa-miR-1-3p were upregulated and the expression levels of hsa-miR-31-5p were downregulated in the experimental group compared with the control group, exhibiting strong original signals and clear differences. .. RT was performed in a 20 µl reaction containing 200 ng total RNA, 0.3 µl 1 µM RT primer, 2 µl 2.5 mM dNTP (HyTest Ltd, Turku, Finland), 2 µl 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µl 50 U/µl RT enzyme (Epicentre; Illumina, Inc.), 0.3 µl 40 U/µl RNase inhibitor (Epicentre; Illumina, Inc.), 20 µl nuclease free water and 0.2 µl MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.). .. The stem-loop RT reaction was performed at 16°C for 30 min, followed by 42°C for 30 min and 85°C for 5 min. Then, RT-qPCR was performed using the SYBR Green I RT-PCR kit (Life Sciences; Thermo Fisher Scientific, Inc.) and GAPDH was used as an internal control, with the following reaction profile: Predenaturation at 95°C for 10 sec, followed by 40 cycles of 95°C for 10 sec and 60°C for 1 min.

    Article Title: MicroRNA profile comparison of the corneal endothelia of young and old mice: implications for senescence of the corneal endothelium
    Article Snippet: Paragraph title: Quantitative reverse transcription polymerase chain reaction microRNA analysis ... Briefly, in a 20 μl reaction volume, 1 μM of each specific reverse transcription primer (0.3 μl), 2 μl dNTP (2.5mM; HyTest Ltd., Turku, Finland), 2 μl 10X RT buffer (Promega Corporation, Madison, WI), 20 U MMLV Reverse Transcriptase (10 U/μl; Epicenter Biotechnologies), 0.3 μL RNase inhibitor (40 U/μl), and 1 μg of total RNA was added. qRT-PCR was performed using the SYBR Green protocol on an ABI 7500 system (Applied Biosystems, Foster City, CA), and a data analysis was performed with the SDS system software (7500 System; Applied Biosystems).

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/ µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/ µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.). .. The stem-loop RT reaction was performed at 16°C for 30 min, followed by 42°C for 30 min and 85°C for 5 min. Then, RT-qPCR was performed using the SYBR Green I RT-PCR kit (Life Sciences; Thermo Fisher Scientific, Inc.) and GAPDH was used as an internal control, with the following reaction profile: Predenaturation at 95°C for 10 sec, followed by 40 cycles of 95°C for 10 sec and 60°C for 1 min.

    Mouse Assay:

    Article Title: MicroRNA profile comparison of the corneal endothelia of young and old mice: implications for senescence of the corneal endothelium
    Article Snippet: Quantitative reverse transcription polymerase chain reaction microRNA analysis To validate the reproducibility of the results from the miRNA microarray, qRT-PCR analysis of (microRNAs come from mice) mmu-miR-695, mmu-miR-183, mmu-miR-182, mmu-miR-194, mmu-miR-34c, mmu-miR-31, mmu-miR-190, and mmu-miR-124 was performed using the same extracted total RNA as the microarray analysis. .. Briefly, in a 20 μl reaction volume, 1 μM of each specific reverse transcription primer (0.3 μl), 2 μl dNTP (2.5mM; HyTest Ltd., Turku, Finland), 2 μl 10X RT buffer (Promega Corporation, Madison, WI), 20 U MMLV Reverse Transcriptase (10 U/μl; Epicenter Biotechnologies), 0.3 μL RNase inhibitor (40 U/μl), and 1 μg of total RNA was added. qRT-PCR was performed using the SYBR Green protocol on an ABI 7500 system (Applied Biosystems, Foster City, CA), and a data analysis was performed with the SDS system software (7500 System; Applied Biosystems).

    Microarray:

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) RT-qPCR analysis was performed to validate selected differently expressed lncRNAs and mRNAs identified by microarray analysis. .. RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Article Title: MicroRNA profile comparison of the corneal endothelia of young and old mice: implications for senescence of the corneal endothelium
    Article Snippet: Quantitative reverse transcription polymerase chain reaction microRNA analysis To validate the reproducibility of the results from the miRNA microarray, qRT-PCR analysis of (microRNAs come from mice) mmu-miR-695, mmu-miR-183, mmu-miR-182, mmu-miR-194, mmu-miR-34c, mmu-miR-31, mmu-miR-190, and mmu-miR-124 was performed using the same extracted total RNA as the microarray analysis. .. Briefly, in a 20 μl reaction volume, 1 μM of each specific reverse transcription primer (0.3 μl), 2 μl dNTP (2.5mM; HyTest Ltd., Turku, Finland), 2 μl 10X RT buffer (Promega Corporation, Madison, WI), 20 U MMLV Reverse Transcriptase (10 U/μl; Epicenter Biotechnologies), 0.3 μL RNase inhibitor (40 U/μl), and 1 μg of total RNA was added. qRT-PCR was performed using the SYBR Green protocol on an ABI 7500 system (Applied Biosystems, Foster City, CA), and a data analysis was performed with the SDS system software (7500 System; Applied Biosystems).

    Article Title: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study
    Article Snippet: RT-qPCR analysis was performed to validate selected differently expressed lncRNAs and mRNAs identified by microarray analysis. .. RT was performed in a 20 µ l reaction containing 200 ng total RNA, 0.3 µ l 1 µ M RT primer, 2 µ l 2.5 mM dNTP (HyTest Ltd., Turku, Finland), 2 µ l 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µ l 50 U/ µ l RT enzyme (Epicentre; Illumina, Inc.), 0.3 µ l 40 U/ µ l RNase inhibitor (Epicentre; Illumina, Inc.), 20 µ l nuclease free water and 0.2 µ l MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Article Title: Differential miRNA expression profiles in human keratinocytes in response to protein kinase C inhibitor
    Article Snippet: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and miRNA target prediction According to the microarray results, the expression levels of hsa-miR-1-3p were upregulated and the expression levels of hsa-miR-31-5p were downregulated in the experimental group compared with the control group, exhibiting strong original signals and clear differences. .. RT was performed in a 20 µl reaction containing 200 ng total RNA, 0.3 µl 1 µM RT primer, 2 µl 2.5 mM dNTP (HyTest Ltd, Turku, Finland), 2 µl 10× RT buffer (Epicentre; Illumina, Inc., San Diego, CA, USA), 1 µl 50 U/µl RT enzyme (Epicentre; Illumina, Inc.), 0.3 µl 40 U/µl RNase inhibitor (Epicentre; Illumina, Inc.), 20 µl nuclease free water and 0.2 µl MMLV High Performance Reverse Transcriptase (Epicentre; Illumina, Inc.).

    Software:

    Article Title: MicroRNA profile comparison of the corneal endothelia of young and old mice: implications for senescence of the corneal endothelium
    Article Snippet: .. Briefly, in a 20 μl reaction volume, 1 μM of each specific reverse transcription primer (0.3 μl), 2 μl dNTP (2.5mM; HyTest Ltd., Turku, Finland), 2 μl 10X RT buffer (Promega Corporation, Madison, WI), 20 U MMLV Reverse Transcriptase (10 U/μl; Epicenter Biotechnologies), 0.3 μL RNase inhibitor (40 U/μl), and 1 μg of total RNA was added. qRT-PCR was performed using the SYBR Green protocol on an ABI 7500 system (Applied Biosystems, Foster City, CA), and a data analysis was performed with the SDS system software (7500 System; Applied Biosystems). .. The levels of an endogenous control, U6 (RiboBio, Guangzhou, China; catalog no. MQP-0201), were used to normalize the expression levels of each miRNA.

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