second strand synthesis  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95

    Structured Review

    Thermo Fisher second strand synthesis
    Second Strand Synthesis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/second strand synthesis/product/Thermo Fisher
    Average 95 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    second strand synthesis - by Bioz Stars, 2020-05
    95/100 stars

    Images

    Related Articles

    Incubation:

    Article Title: Type I and type II interferon responses in two human liver cell lines (Huh-7 and HuH6)
    Article Snippet: .. Next, the first-strand buffer mix (4 μL of 5 × the first-strand buffer, 2 μL 0.1 M dithiothreitol (DTT), and 1 μL 10 mM dNTPs) was preincubated at 42 °C for 2 min. After addition of 2 μL (200 units) Superscript II (Life Technologies, Karlsruhe, Germany), incubation was continued at 42 °C for 1 h. For second-strand synthesis, 30 μL 5 × the second-strand buffer, 91 μL RNase-free water, 3 μL 10 mM dNTPs, 4 μL (40 U) Escherichia coli DNA polymerase I (Life Technologies), 1 μL (12 U) E. coli DNAligase (TaKaRa, Gennevilliers, France), and 1 μL (2 U) RNase H (TaKaRa) were added, and the mix was incubated at 16 °C for 2 h. Then 2.5 μL (10 U) T4 DNA polymerase I (TaKaRa) were added at 16 °C for 5 min. .. The reaction was stopped by the addition of 10 μL 0.5 M EDTA, double-stranded (ds) cDNA was extracted with phenol/chloroform, and the aqueous phase was recovered by phase-lock gel separation (Eppendorf, Hamburg, Germany).

    Article Title: Genome Wide Association Identifies Novel Loci Involved in Fungal Communication
    Article Snippet: .. For second strand synthesis, 51 µL of H2 O, 20 µL of 5× second strand buffer (Invitrogen 10812-014), and dNTPs (10 mM) were added to the first strand cDNA synthesis mix and incubated on ice for 5 minutes. .. RNaseH (2 U) (Invitrogen 18021-014), DNA pol I (50 U) (Invitrogen 18010-017) were then added and the mixture was incubated at 16°C for 2.5 hours.

    Article Title: Global DNA compaction in stationary-phase bacteria does not affect transcription
    Article Snippet: .. To synthesize cDNA, 1 μL 50 μM Random hexamers (Invitrogen), 1 μg total RNA, 1 μL 10 mM dNTP mix (Promega), and nuclease-free water (Promega) up to 13 μL were mixed and subsequently heated to 65°C for 5 min and then cooled on ice for at least 1 min. After cooling, 4 μL 5× first strand buffer (Invitrogen), 1 μL 0.1M DTT, 1 μL RNasin® RNase inhibitor (Promega) and 1μL of SuperScript III Reverse Transcriptase (Invitrogen) was added and mixtures were incubated at 25°C for 5 min, at 50°C for 1 h and then at 70°C for 15 min. After the first strand synthesis, the following components were added; 30 μL second strand buffer (Invitrogen), 3 μL of 10mM dNTP mix (Promega), 4 μL of E. coli DNA polymerase I (NEB), 1 μL E. coli DNA ligase (New England Biolabs), 1 μL 5 U/μL RNase H, and 91 μL nuclease-free water. .. Second strand synthesis mixtures were incubated at 16°C for 2 h. The resulting double-stranded cDNA was then purified with the SV DNA purification kit from Promega.

    Article Title: Genetic basis of transcriptome diversity in Drosophila melanogaster
    Article Snippet: .. For each of the two replicates for each line and each sex, first-strand cDNA was prepared from 7 μg of total RNA (1 μg/μL) with 1 μL of random primers (3 μg/μL) (Invitrogen) and incubation at 70 °C for 5 min, 25 ° C for 5 min, and 4 °C for 10 min. We added 5× first-strand buffer (4 μL) (Invitrogen), 0.1 M DTT (2 μL) (Invitrogen), 10 mM dNTP+dUTP (1 μL) (Promega), RNase Inhibitor (1 μL) (Invitrogen), and SuperScript II (4 μL) (Invitrogen) and incubated the reactions in a thermal cycler (with a heated lid) using the following program: 25 °C for 10 min; 42 °C for 90 min; 70 °C for 10 min; and 4 °C for 10 min. Second-strand cDNA was synthesized by adding 17.5 mM MgCl2 (8 μL) (Sigma), 10 mM dNTP+dUTP (1 μL) (Promega), DNA Polymerase I (1.2 μL) (Promega), RNase H (0.5 μL) (Promega), and RNase-free water (9.3 μL) (Ambion) to the first-strand cDNA reactions. .. The reactions were incubated in a thermal cycler at 16 °C for 2 h (without a heated lid) followed by 75 °C for 10 min (with a heated lid) and 4 °C for 10 min. Double-stranded cDNA was purified using the QIAquick 96 PCR kit (Qiagen) by following the manufacturer’s protocol except that buffer PN (Qiagen) was used instead of buffer PM (Qiagen).

    Synthesized:

    Article Title: Genome-wide p63-regulated gene expression in differentiating epidermal keratinocytes
    Article Snippet: .. The second strand was synthesized by adding to the purified sample (100 μl total volume): 20 μl 5 × Second strand buffer (Invitrogen), 4 μl 5 × First strand buffer (Invitrogen), 2 μl DTT (0.1 M Invitrogen), 1 μl random hexamers (5 mg/ml Roche), dUTP mix (12.5 mM Invitrogen), 1 μl RNase H (8 U/ml Ambion), 1 μl E scherichia coli DNA polymerase I (10 U/μl Invitrogen), and 1 μl E. coli DNA ligase (10 U/μl NEB). .. After 2 h at 16 °C, 1 μl of T4 DNA polymerase (10 U/μl Promega) was added and the reaction was incubated at 16 °C for 10 min.

    Article Title: Genetic basis of transcriptome diversity in Drosophila melanogaster
    Article Snippet: .. For each of the two replicates for each line and each sex, first-strand cDNA was prepared from 7 μg of total RNA (1 μg/μL) with 1 μL of random primers (3 μg/μL) (Invitrogen) and incubation at 70 °C for 5 min, 25 ° C for 5 min, and 4 °C for 10 min. We added 5× first-strand buffer (4 μL) (Invitrogen), 0.1 M DTT (2 μL) (Invitrogen), 10 mM dNTP+dUTP (1 μL) (Promega), RNase Inhibitor (1 μL) (Invitrogen), and SuperScript II (4 μL) (Invitrogen) and incubated the reactions in a thermal cycler (with a heated lid) using the following program: 25 °C for 10 min; 42 °C for 90 min; 70 °C for 10 min; and 4 °C for 10 min. Second-strand cDNA was synthesized by adding 17.5 mM MgCl2 (8 μL) (Sigma), 10 mM dNTP+dUTP (1 μL) (Promega), DNA Polymerase I (1.2 μL) (Promega), RNase H (0.5 μL) (Promega), and RNase-free water (9.3 μL) (Ambion) to the first-strand cDNA reactions. .. The reactions were incubated in a thermal cycler at 16 °C for 2 h (without a heated lid) followed by 75 °C for 10 min (with a heated lid) and 4 °C for 10 min. Double-stranded cDNA was purified using the QIAquick 96 PCR kit (Qiagen) by following the manufacturer’s protocol except that buffer PN (Qiagen) was used instead of buffer PM (Qiagen).

    Purification:

    Article Title: Genome-wide p63-regulated gene expression in differentiating epidermal keratinocytes
    Article Snippet: .. The second strand was synthesized by adding to the purified sample (100 μl total volume): 20 μl 5 × Second strand buffer (Invitrogen), 4 μl 5 × First strand buffer (Invitrogen), 2 μl DTT (0.1 M Invitrogen), 1 μl random hexamers (5 mg/ml Roche), dUTP mix (12.5 mM Invitrogen), 1 μl RNase H (8 U/ml Ambion), 1 μl E scherichia coli DNA polymerase I (10 U/μl Invitrogen), and 1 μl E. coli DNA ligase (10 U/μl NEB). .. After 2 h at 16 °C, 1 μl of T4 DNA polymerase (10 U/μl Promega) was added and the reaction was incubated at 16 °C for 10 min.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 98
    Thermo Fisher revertaid premium
    Expression in breast cancer cell lines of KAI1 as-lncRNA, KAI1 mRNA, and comparison of their in vitro invasion potency (A) MCF-7, MDA-MB-231, MDA-MB-435 and SUM149PT cell lines derived RNAs were subjected to DNase. Then 1μg RNA of each was reverse transcribed with <t>RevertAid</t> Premium followed by quantitative Real-Time PCR method. (B) 1μg RNA of each of the cell lines was reverse transcribed using oligodT (15) priming. KAI1 mRNA levels relative to endogenous control HPRT-1 mRNA were evaluated by quantitative Real-Time PCR. (C) The four indicated cell lines were seeded in Matrigel coated transwell chambers. Cells invading from upper chamber to the chemo attractant (10% serum) in the lower chamber were detected using resazurin cell viability assay. Images of random fields were taken by light-field microscope camera as illustrative support.
    Revertaid Premium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/revertaid premium/product/Thermo Fisher
    Average 98 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    revertaid premium - by Bioz Stars, 2020-05
    98/100 stars
      Buy from Supplier

    93
    Thermo Fisher dna polymerase i
    Evaluation of the HexaPrime assay. (A) Evaluation of different primer pairs for the detection of coronaviruses. Analysis was conducted using the HCoV-NL63 virus and all primer sets given in Table 2 were tested. Only amplification with primer sets 2, 4, 5 and 8 yielded distinct bands. Sequencing of products and analysis of fragment size revealed that only primer set 2 allowed efficient amplification of the desired product. M: size marker; mock-infected (−) or HCoV-NL63-infected (+) cell culture supernatant. (B) Detection of HCoV-NL63 and HCoV-HKU1 with the HexaPrime assay using primer set 2. All experimental procedures were conducted as described in Section 2 . M: size marker; W: water; NL63 and HKU1: mock-infected (−) or virus-infected (+) cell culture supernatant. (C) Sensitivity of the HexaPrime assay. Concentrated samples containing viral RNA (10 9 copies ml −1 ) were subjected to 10-fold serial dilutions in cell culture supernatant and the HexaPrime assay was conducted. For each RNA concentration, three different enzymes for SS <t>DNA</t> synthesis were trialed. A, B and C denote DNA Polymerase I, T7 Polymerase, and Sequenase 2.0, respectively.
    Dna Polymerase I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna polymerase i/product/Thermo Fisher
    Average 93 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    dna polymerase i - by Bioz Stars, 2020-05
    93/100 stars
      Buy from Supplier

    99
    Thermo Fisher second strand cdna synthesis
    Evaluation of the HexaPrime assay. (A) Evaluation of different primer pairs for the detection of coronaviruses. Analysis was conducted using the HCoV-NL63 virus and all primer sets given in Table 2 were tested. Only amplification with primer sets 2, 4, 5 and 8 yielded distinct bands. Sequencing of products and analysis of fragment size revealed that only primer set 2 allowed efficient amplification of the desired product. M: size marker; mock-infected (−) or HCoV-NL63-infected (+) cell culture supernatant. (B) Detection of HCoV-NL63 and HCoV-HKU1 with the HexaPrime assay using primer set 2. All experimental procedures were conducted as described in Section 2 . M: size marker; W: water; NL63 and HKU1: mock-infected (−) or virus-infected (+) cell culture supernatant. (C) Sensitivity of the HexaPrime assay. Concentrated samples containing viral RNA (10 9 copies ml −1 ) were subjected to 10-fold serial dilutions in cell culture supernatant and the HexaPrime assay was conducted. For each RNA concentration, three different enzymes for SS <t>DNA</t> synthesis were trialed. A, B and C denote DNA Polymerase I, T7 Polymerase, and Sequenase 2.0, respectively.
    Second Strand Cdna Synthesis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/second strand cdna synthesis/product/Thermo Fisher
    Average 99 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    second strand cdna synthesis - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    Image Search Results


    Expression in breast cancer cell lines of KAI1 as-lncRNA, KAI1 mRNA, and comparison of their in vitro invasion potency (A) MCF-7, MDA-MB-231, MDA-MB-435 and SUM149PT cell lines derived RNAs were subjected to DNase. Then 1μg RNA of each was reverse transcribed with RevertAid Premium followed by quantitative Real-Time PCR method. (B) 1μg RNA of each of the cell lines was reverse transcribed using oligodT (15) priming. KAI1 mRNA levels relative to endogenous control HPRT-1 mRNA were evaluated by quantitative Real-Time PCR. (C) The four indicated cell lines were seeded in Matrigel coated transwell chambers. Cells invading from upper chamber to the chemo attractant (10% serum) in the lower chamber were detected using resazurin cell viability assay. Images of random fields were taken by light-field microscope camera as illustrative support.

    Journal: Oncotarget

    Article Title: Identification of a novel metastasis inducing lncRNA which suppresses the KAI1/CD82 metastasis suppressor gene and is upregulated in triple-negative breast cancer

    doi: 10.18632/oncotarget.18733

    Figure Lengend Snippet: Expression in breast cancer cell lines of KAI1 as-lncRNA, KAI1 mRNA, and comparison of their in vitro invasion potency (A) MCF-7, MDA-MB-231, MDA-MB-435 and SUM149PT cell lines derived RNAs were subjected to DNase. Then 1μg RNA of each was reverse transcribed with RevertAid Premium followed by quantitative Real-Time PCR method. (B) 1μg RNA of each of the cell lines was reverse transcribed using oligodT (15) priming. KAI1 mRNA levels relative to endogenous control HPRT-1 mRNA were evaluated by quantitative Real-Time PCR. (C) The four indicated cell lines were seeded in Matrigel coated transwell chambers. Cells invading from upper chamber to the chemo attractant (10% serum) in the lower chamber were detected using resazurin cell viability assay. Images of random fields were taken by light-field microscope camera as illustrative support.

    Article Snippet: Second strand synthesis with RevertAid Premium (Thermo Scientific) was performed using an adapter with a polyT tail to prime the generated polyA tail of the cDNA.

    Techniques: Expressing, In Vitro, Multiple Displacement Amplification, Derivative Assay, Real-time Polymerase Chain Reaction, Viability Assay, Microscopy

    Quantitation of KAI1 as-lncRNA following its shRNA mediated knockdown in MDA-MB-231, MDA-MB-435 and MCF-7 cells qRT-PCR after MDA-MB-435 (A) , MDA-MB-231 (B) and MCF-7 cells (C) infection with SHC-shRNA vector empty, SHC-shRNA- non-silencing (ns) or shRNA-1 against KAI1 as-lncRNA (SHC-shRNA-1). All samples were reverse transcribed with RevertAid Premium RTase (or without RTase as control). Results are average of five independent experiments in triplicates. HPRT-1 mRNA served as endogenous control. The mean of sh-ns was set to 1 in panels (A C), with the mean value of sh-empty set to 1 in panel (B). p (“SHC-shRNA-1”)

    Journal: Oncotarget

    Article Title: Identification of a novel metastasis inducing lncRNA which suppresses the KAI1/CD82 metastasis suppressor gene and is upregulated in triple-negative breast cancer

    doi: 10.18632/oncotarget.18733

    Figure Lengend Snippet: Quantitation of KAI1 as-lncRNA following its shRNA mediated knockdown in MDA-MB-231, MDA-MB-435 and MCF-7 cells qRT-PCR after MDA-MB-435 (A) , MDA-MB-231 (B) and MCF-7 cells (C) infection with SHC-shRNA vector empty, SHC-shRNA- non-silencing (ns) or shRNA-1 against KAI1 as-lncRNA (SHC-shRNA-1). All samples were reverse transcribed with RevertAid Premium RTase (or without RTase as control). Results are average of five independent experiments in triplicates. HPRT-1 mRNA served as endogenous control. The mean of sh-ns was set to 1 in panels (A C), with the mean value of sh-empty set to 1 in panel (B). p (“SHC-shRNA-1”)

    Article Snippet: Second strand synthesis with RevertAid Premium (Thermo Scientific) was performed using an adapter with a polyT tail to prime the generated polyA tail of the cDNA.

    Techniques: Quantitation Assay, shRNA, Multiple Displacement Amplification, Quantitative RT-PCR, Infection, Plasmid Preparation

    Evaluation of the HexaPrime assay. (A) Evaluation of different primer pairs for the detection of coronaviruses. Analysis was conducted using the HCoV-NL63 virus and all primer sets given in Table 2 were tested. Only amplification with primer sets 2, 4, 5 and 8 yielded distinct bands. Sequencing of products and analysis of fragment size revealed that only primer set 2 allowed efficient amplification of the desired product. M: size marker; mock-infected (−) or HCoV-NL63-infected (+) cell culture supernatant. (B) Detection of HCoV-NL63 and HCoV-HKU1 with the HexaPrime assay using primer set 2. All experimental procedures were conducted as described in Section 2 . M: size marker; W: water; NL63 and HKU1: mock-infected (−) or virus-infected (+) cell culture supernatant. (C) Sensitivity of the HexaPrime assay. Concentrated samples containing viral RNA (10 9 copies ml −1 ) were subjected to 10-fold serial dilutions in cell culture supernatant and the HexaPrime assay was conducted. For each RNA concentration, three different enzymes for SS DNA synthesis were trialed. A, B and C denote DNA Polymerase I, T7 Polymerase, and Sequenase 2.0, respectively.

    Journal: Journal of Virological Methods

    Article Title: HexaPrime: A novel method for detection of coronaviruses

    doi: 10.1016/j.jviromet.2012.11.039

    Figure Lengend Snippet: Evaluation of the HexaPrime assay. (A) Evaluation of different primer pairs for the detection of coronaviruses. Analysis was conducted using the HCoV-NL63 virus and all primer sets given in Table 2 were tested. Only amplification with primer sets 2, 4, 5 and 8 yielded distinct bands. Sequencing of products and analysis of fragment size revealed that only primer set 2 allowed efficient amplification of the desired product. M: size marker; mock-infected (−) or HCoV-NL63-infected (+) cell culture supernatant. (B) Detection of HCoV-NL63 and HCoV-HKU1 with the HexaPrime assay using primer set 2. All experimental procedures were conducted as described in Section 2 . M: size marker; W: water; NL63 and HKU1: mock-infected (−) or virus-infected (+) cell culture supernatant. (C) Sensitivity of the HexaPrime assay. Concentrated samples containing viral RNA (10 9 copies ml −1 ) were subjected to 10-fold serial dilutions in cell culture supernatant and the HexaPrime assay was conducted. For each RNA concentration, three different enzymes for SS DNA synthesis were trialed. A, B and C denote DNA Polymerase I, T7 Polymerase, and Sequenase 2.0, respectively.

    Article Snippet: Second-strand synthesis was conducted using DNA Polymerase I and the SS primers listed in .

    Techniques: Amplification, Sequencing, Marker, Infection, Cell Culture, Concentration Assay, DNA Synthesis