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    Thermo Fisher reverse transcriptase superscript ii
    ( a ) Results of qRT-PCR showing (i) Brn-3a and (ii) Brn-3b mRNA levels in the developing zebrafish at 24, 48 and 72 hpf. <t>cDNA</t> from total <t>RNA</t> was amplified using primers to ZF Brn-3a and Brn-3b and a variation in mRNA levels was corrected using ZF GAPDH. Values were expressed as fold induction relative to expression at 24 h (set at 1). ( b ) Representative western blot analysis showing single protein band for both Brn-3a and Brn-3b in extracts from adult ZF compared with adult mouse heart (M), used as a positive control. MW markers indicate the protein size and gamma ( γ ) tubulin was used to control for variation in total protein. ( c ) Representative images showing whole-mount immunostaining for (i) Brn-3a or (ii) Brn-3b (green; top panels) in ZF hearts at 72 hpf. Co-staining with tropomyosin (red; middle panels) indicate cardiomyocytes in the developing heart. Lower panel shows merge with bright field image. V, A, P (indicated by arrow). ( d ) Representative images of DAB-immunostained ZF embryos sections at 72 hpf. Protein localisation is seen as dark brown staining in ventricles (indicated by arrow), identified by TM, also shows Brn-3a and Brn-3b expression. × 40 magnification. A, atria; hpf, hours post fertilisation; MW, molecular weight; P, pericardial sac; TM, tropomyosin; V, ventricle; ZF, zebrafish heart
    Reverse Transcriptase Superscript Ii, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8919 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reverse transcriptase superscript ii/product/Thermo Fisher
    Average 99 stars, based on 8919 article reviews
    Price from $9.99 to $1999.99
    reverse transcriptase superscript ii - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher superscript ii rna reverse transcriptase
    ( a ) Results of qRT-PCR showing (i) Brn-3a and (ii) Brn-3b mRNA levels in the developing zebrafish at 24, 48 and 72 hpf. <t>cDNA</t> from total <t>RNA</t> was amplified using primers to ZF Brn-3a and Brn-3b and a variation in mRNA levels was corrected using ZF GAPDH. Values were expressed as fold induction relative to expression at 24 h (set at 1). ( b ) Representative western blot analysis showing single protein band for both Brn-3a and Brn-3b in extracts from adult ZF compared with adult mouse heart (M), used as a positive control. MW markers indicate the protein size and gamma ( γ ) tubulin was used to control for variation in total protein. ( c ) Representative images showing whole-mount immunostaining for (i) Brn-3a or (ii) Brn-3b (green; top panels) in ZF hearts at 72 hpf. Co-staining with tropomyosin (red; middle panels) indicate cardiomyocytes in the developing heart. Lower panel shows merge with bright field image. V, A, P (indicated by arrow). ( d ) Representative images of DAB-immunostained ZF embryos sections at 72 hpf. Protein localisation is seen as dark brown staining in ventricles (indicated by arrow), identified by TM, also shows Brn-3a and Brn-3b expression. × 40 magnification. A, atria; hpf, hours post fertilisation; MW, molecular weight; P, pericardial sac; TM, tropomyosin; V, ventricle; ZF, zebrafish heart
    Superscript Ii Rna Reverse Transcriptase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript ii rna reverse transcriptase/product/Thermo Fisher
    Average 99 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    superscript ii rna reverse transcriptase - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    Image Search Results


    ( a ) Results of qRT-PCR showing (i) Brn-3a and (ii) Brn-3b mRNA levels in the developing zebrafish at 24, 48 and 72 hpf. cDNA from total RNA was amplified using primers to ZF Brn-3a and Brn-3b and a variation in mRNA levels was corrected using ZF GAPDH. Values were expressed as fold induction relative to expression at 24 h (set at 1). ( b ) Representative western blot analysis showing single protein band for both Brn-3a and Brn-3b in extracts from adult ZF compared with adult mouse heart (M), used as a positive control. MW markers indicate the protein size and gamma ( γ ) tubulin was used to control for variation in total protein. ( c ) Representative images showing whole-mount immunostaining for (i) Brn-3a or (ii) Brn-3b (green; top panels) in ZF hearts at 72 hpf. Co-staining with tropomyosin (red; middle panels) indicate cardiomyocytes in the developing heart. Lower panel shows merge with bright field image. V, A, P (indicated by arrow). ( d ) Representative images of DAB-immunostained ZF embryos sections at 72 hpf. Protein localisation is seen as dark brown staining in ventricles (indicated by arrow), identified by TM, also shows Brn-3a and Brn-3b expression. × 40 magnification. A, atria; hpf, hours post fertilisation; MW, molecular weight; P, pericardial sac; TM, tropomyosin; V, ventricle; ZF, zebrafish heart

    Journal: Cell Death & Disease

    Article Title: Essential but partially redundant roles for POU4F1/Brn-3a and POU4F2/Brn-3b transcription factors in the developing heart

    doi: 10.1038/cddis.2017.185

    Figure Lengend Snippet: ( a ) Results of qRT-PCR showing (i) Brn-3a and (ii) Brn-3b mRNA levels in the developing zebrafish at 24, 48 and 72 hpf. cDNA from total RNA was amplified using primers to ZF Brn-3a and Brn-3b and a variation in mRNA levels was corrected using ZF GAPDH. Values were expressed as fold induction relative to expression at 24 h (set at 1). ( b ) Representative western blot analysis showing single protein band for both Brn-3a and Brn-3b in extracts from adult ZF compared with adult mouse heart (M), used as a positive control. MW markers indicate the protein size and gamma ( γ ) tubulin was used to control for variation in total protein. ( c ) Representative images showing whole-mount immunostaining for (i) Brn-3a or (ii) Brn-3b (green; top panels) in ZF hearts at 72 hpf. Co-staining with tropomyosin (red; middle panels) indicate cardiomyocytes in the developing heart. Lower panel shows merge with bright field image. V, A, P (indicated by arrow). ( d ) Representative images of DAB-immunostained ZF embryos sections at 72 hpf. Protein localisation is seen as dark brown staining in ventricles (indicated by arrow), identified by TM, also shows Brn-3a and Brn-3b expression. × 40 magnification. A, atria; hpf, hours post fertilisation; MW, molecular weight; P, pericardial sac; TM, tropomyosin; V, ventricle; ZF, zebrafish heart

    Article Snippet: RNA was quantified (NanoDrop 1000 spectrophotometer, Thermo Fisher Scientific, Paisley, UK) and cDNA synthesis (20–50 μ l reaction) was carried out using RNA Superscript II Reverse Transcriptase (Invitrogen).

    Techniques: Quantitative RT-PCR, Amplification, Expressing, Western Blot, Positive Control, Immunostaining, Staining, Molecular Weight

    Full-length L1Rn elements are transcribed in RCL cells. ( A ) Structure of L1Rn. A functional full-length L1Rn element is characterized by two ORFs flanked by 5′ and 3′ UTRs. The bipartite 5′ UTR consists of a monomer, which can be tandemly repeated, and a non-repeated tether (t). The most 5′ monomer is only partially duplicated (black truncated box with arrowhead) in all genomic rat elements identified so far. Horizontal arrows indicate the binding sites of the oligonucleotides Pr-L1Rn1 and Pr-L1Rn2 used to amplify cDNAs generated from full-length transcripts. The binding site of the 500-bp digoxigenin-labeled probe used to detect L1-specific transcripts is localized at the 5′ end of ORF2 (black bar). Open bars represent ORF1- and ORF2-encoded polypeptides against which monoclonal antibodies were raised. The polypeptides are covering amino acid positions 144–402 and 292–480 of ORF1p and ORF2p, respectively (accession no. DQ100480). LPR, length polymorphism region; GrPPT, G-rich polypurine tract; A n , A-rich 3′ tract; EN, endonuclease; RT, reverse transcriptase; C, cysteine-rich motif. ( B ) L1Rn transcriptional products in RCL cells. PolyA + RNA was isolated from RCL cells that had reached the maximum population density of ∼10 6 cells/ml (lane 2) and from cells that were UV-irradiated (lane 1). Two microgram of each RNA were separated by agarose gel electrophoresis and subjected to northern blot analysis using a 500-bp probe ( Figure 1 A). (C) Schematic structures of 10 L1Rn cDNAs synthesized from poly(A) + RNA from UV-irradiated RCL cells. cDNAs are flanked by primer sequences Pr-L1Rn1 and Pr-L1Rn2. Names of the resulting cDNAs are listed on the left, while accession numbers are specified on the right. Termination codons within ORF2 sequences are indicated by vertical arrows. Two deletions in the ORF2-coding region of cDNA L1-17 covering 103 and 220 nts, respectively, are indicated by interrupted bars.

    Journal: Nucleic Acids Research

    Article Title: Functional endogenous LINE-1 retrotransposons are expressed and mobilized in rat chloroleukemia cells

    doi: 10.1093/nar/gkm1045

    Figure Lengend Snippet: Full-length L1Rn elements are transcribed in RCL cells. ( A ) Structure of L1Rn. A functional full-length L1Rn element is characterized by two ORFs flanked by 5′ and 3′ UTRs. The bipartite 5′ UTR consists of a monomer, which can be tandemly repeated, and a non-repeated tether (t). The most 5′ monomer is only partially duplicated (black truncated box with arrowhead) in all genomic rat elements identified so far. Horizontal arrows indicate the binding sites of the oligonucleotides Pr-L1Rn1 and Pr-L1Rn2 used to amplify cDNAs generated from full-length transcripts. The binding site of the 500-bp digoxigenin-labeled probe used to detect L1-specific transcripts is localized at the 5′ end of ORF2 (black bar). Open bars represent ORF1- and ORF2-encoded polypeptides against which monoclonal antibodies were raised. The polypeptides are covering amino acid positions 144–402 and 292–480 of ORF1p and ORF2p, respectively (accession no. DQ100480). LPR, length polymorphism region; GrPPT, G-rich polypurine tract; A n , A-rich 3′ tract; EN, endonuclease; RT, reverse transcriptase; C, cysteine-rich motif. ( B ) L1Rn transcriptional products in RCL cells. PolyA + RNA was isolated from RCL cells that had reached the maximum population density of ∼10 6 cells/ml (lane 2) and from cells that were UV-irradiated (lane 1). Two microgram of each RNA were separated by agarose gel electrophoresis and subjected to northern blot analysis using a 500-bp probe ( Figure 1 A). (C) Schematic structures of 10 L1Rn cDNAs synthesized from poly(A) + RNA from UV-irradiated RCL cells. cDNAs are flanked by primer sequences Pr-L1Rn1 and Pr-L1Rn2. Names of the resulting cDNAs are listed on the left, while accession numbers are specified on the right. Termination codons within ORF2 sequences are indicated by vertical arrows. Two deletions in the ORF2-coding region of cDNA L1-17 covering 103 and 220 nts, respectively, are indicated by interrupted bars.

    Article Snippet: Cloning of transcribed L1Rn sequences from RCL cells Total and poly(A)+ RNA were prepared from control and irradiated RCL cells with the RNeasy and Oligotex mRNA kits (Qiagen), respectively. cDNA was synthesized from poly(A)+ RNA employing SuperScript™ II reverse transcriptase (Invitrogen) and oligo(dT) primer.

    Techniques: Functional Assay, Binding Assay, Generated, Labeling, Isolation, Irradiation, Agarose Gel Electrophoresis, Northern Blot, Synthesized

    Deep sequencing analyses of control linear RNAs and RNAs from young (4-month-old) and old (2-year-old) brain samples ( A ) Similar performance of RNA-Seq analyses on a library of 96 linear RNAs (ERCC RNA Spike-In Mix) using SuperScript II and MonsterScript. Data represent mean values of duplicated experiments. ( B ) Twenty artificial circular RNAs were identified from SuperScript II, but none from MonsterScript libraries of the ERCC RNA Spike-In Mix based on junction sequences. ( C ) Relationship between sequencing depth and noise-to-signal ratios of SuperScript II vs. MonsterScript-based library construction protocols for circular RNA identification. Sequencing data of the ERCC RNA Spike-In Mix were used for this analysis. ( D ) The scatter plot showing sequencing count distribution of all the genes detected in the duplicated libraries constructed using the two reverse transcriptases (MonsterScript and SuperScript II) using RNAs from brains samples of young (4-month-old) and old (2-year-old) mice. ( E ) Venn diagram showing the putative circular RNAs identified from mouse brain by a previous ( 8 ) and the present studies using SuperScript II- and MonsterScript-based library construction protocols. Junction reads ratios in mouse brain libraries from a previous and the present studies using SuperScript II and MonsterScript-based library construction protocols. ( F ) Potential motifs found in the junction sequences of artificial circular RNAs.

    Journal: bioRxiv

    Article Title: Template switching causes artificial junction formation and false identification of circular RNAs

    doi: 10.1101/259556

    Figure Lengend Snippet: Deep sequencing analyses of control linear RNAs and RNAs from young (4-month-old) and old (2-year-old) brain samples ( A ) Similar performance of RNA-Seq analyses on a library of 96 linear RNAs (ERCC RNA Spike-In Mix) using SuperScript II and MonsterScript. Data represent mean values of duplicated experiments. ( B ) Twenty artificial circular RNAs were identified from SuperScript II, but none from MonsterScript libraries of the ERCC RNA Spike-In Mix based on junction sequences. ( C ) Relationship between sequencing depth and noise-to-signal ratios of SuperScript II vs. MonsterScript-based library construction protocols for circular RNA identification. Sequencing data of the ERCC RNA Spike-In Mix were used for this analysis. ( D ) The scatter plot showing sequencing count distribution of all the genes detected in the duplicated libraries constructed using the two reverse transcriptases (MonsterScript and SuperScript II) using RNAs from brains samples of young (4-month-old) and old (2-year-old) mice. ( E ) Venn diagram showing the putative circular RNAs identified from mouse brain by a previous ( 8 ) and the present studies using SuperScript II- and MonsterScript-based library construction protocols. Junction reads ratios in mouse brain libraries from a previous and the present studies using SuperScript II and MonsterScript-based library construction protocols. ( F ) Potential motifs found in the junction sequences of artificial circular RNAs.

    Article Snippet: RT-PCR analyses of linear and circular RNAs Cellular RNAs or synthetic control linear or circular RNAs were reverse transcribed using MonsterScript (Cat# MS041050, Epicentre) and SuperScript II (Cat# 18064014, Thermo Fisher) in a reaction containing the following reagents: 4μl 5xBuffer (supplementing 10mM DTT for SuperScript II), 1μl dNTP, 1μl reverse transcriptase, 1μl control RNA, 1μl Smarter IIA oligo (10μM), 1μl gene-specific primer (10uM), 11μl water.

    Techniques: Sequencing, RNA Sequencing Assay, Construct, Mouse Assay

    Junction formation due to template-switching (TS) when commonly used reverse transcriptases (e.g., SuperScript II) are used for cDNA synthesis ( A ) Schematic presentation of the mechanism through which the terminal transferase activity of MMLV-derived reverse transcriptases (e.g., SuperScript II) can add several non-templated nucleotides to the 3’ end of cDNA, leading to TS during reverse transcription (RT). The TS events can generate artificial junctions derived from the same linear RNA templates and these linear RNAs would be erroneously identified as RNA circles in bioinformatic analyses. ( B ) PCR detection of the junction sequences from SuperScript II or MonsterScript RT products of both control circular (sample 1) and linear (sample 2) RNAs using convergent and divergent primers, respectively (upper left half panel). PCRs using internal primers were used as loading controls (upper right half panel). The lower panels show results after RNase R treatment of the control linear and circular RNAs. ( C ) Sanger sequencing of the PCR products. Note that the divergent PCR products of SuperScript II cDNAs from the control linear RNA contained insertions and mutations resulting from TS events, whereas both SuperScript II and MonsterScript convergent PCR products were from the true RNA ligation junctions of the control circular RNA. Fluc, control luciferase RNA; MS, MonsterScript; SSI I: SuperScript II. ( D ) Reverse transcriptase efficiency assays using the control linear RNA (left panel) and a previously validated circular RNA, Cdr1as (right panel). RNAs were reverse transcribed using SuperScript II and MonsterScript, followed by qPCR using the internal primers (for control linear RNA) and junction primers (for Cdr1as), respectively. CT values were plotted against log 10 values of RNA inputs and SuperScript II and MonsterScript displayed similar sensitivity in detecting either linear or circular RNAs.

    Journal: bioRxiv

    Article Title: Template switching causes artificial junction formation and false identification of circular RNAs

    doi: 10.1101/259556

    Figure Lengend Snippet: Junction formation due to template-switching (TS) when commonly used reverse transcriptases (e.g., SuperScript II) are used for cDNA synthesis ( A ) Schematic presentation of the mechanism through which the terminal transferase activity of MMLV-derived reverse transcriptases (e.g., SuperScript II) can add several non-templated nucleotides to the 3’ end of cDNA, leading to TS during reverse transcription (RT). The TS events can generate artificial junctions derived from the same linear RNA templates and these linear RNAs would be erroneously identified as RNA circles in bioinformatic analyses. ( B ) PCR detection of the junction sequences from SuperScript II or MonsterScript RT products of both control circular (sample 1) and linear (sample 2) RNAs using convergent and divergent primers, respectively (upper left half panel). PCRs using internal primers were used as loading controls (upper right half panel). The lower panels show results after RNase R treatment of the control linear and circular RNAs. ( C ) Sanger sequencing of the PCR products. Note that the divergent PCR products of SuperScript II cDNAs from the control linear RNA contained insertions and mutations resulting from TS events, whereas both SuperScript II and MonsterScript convergent PCR products were from the true RNA ligation junctions of the control circular RNA. Fluc, control luciferase RNA; MS, MonsterScript; SSI I: SuperScript II. ( D ) Reverse transcriptase efficiency assays using the control linear RNA (left panel) and a previously validated circular RNA, Cdr1as (right panel). RNAs were reverse transcribed using SuperScript II and MonsterScript, followed by qPCR using the internal primers (for control linear RNA) and junction primers (for Cdr1as), respectively. CT values were plotted against log 10 values of RNA inputs and SuperScript II and MonsterScript displayed similar sensitivity in detecting either linear or circular RNAs.

    Article Snippet: RT-PCR analyses of linear and circular RNAs Cellular RNAs or synthetic control linear or circular RNAs were reverse transcribed using MonsterScript (Cat# MS041050, Epicentre) and SuperScript II (Cat# 18064014, Thermo Fisher) in a reaction containing the following reagents: 4μl 5xBuffer (supplementing 10mM DTT for SuperScript II), 1μl dNTP, 1μl reverse transcriptase, 1μl control RNA, 1μl Smarter IIA oligo (10μM), 1μl gene-specific primer (10uM), 11μl water.

    Techniques: Activity Assay, Derivative Assay, Polymerase Chain Reaction, Sequencing, Ligation, Luciferase, Real-time Polymerase Chain Reaction