rtcb ligase  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Name:
    RtcB Ligase
    Description:

    Catalog Number:
    M0458
    Price:
    74
    Category:
    Enzymes for Innovation
    Applications:
    Functional Genomics
    Size:
    25 reactions
    Buy from Supplier


    Structured Review

    New England Biolabs rtcb ligase
    RtcB Ligase

    https://www.bioz.com/result/rtcb ligase/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rtcb ligase - by Bioz Stars, 2021-07
    94/100 stars

    Images

    1) Product Images from "Small circRNAs with self-cleaving ribozymes are frequently expressed in metazoan transcriptomes"

    Article Title: Small circRNAs with self-cleaving ribozymes are frequently expressed in metazoan transcriptomes

    Journal: bioRxiv

    doi: 10.1101/721605

    Circularization of Mexican axolotl self-cleaved retrozyme RNA. Purified linear monomeric RNA resulting from double self-cleavage of DRtz353 (see Figure 4C ) was run directly in a denaturing gel (lane 1), after 1h incubation with RtcB ligase in its corresponding buffer (lane 2), in RtcB buffer without protein ligase (lane 3), and in 50 mM Mg 2+ buffer (lane 4). Circular RNAs can be readily detected in lane 2 (up to 80% circularization for the 350 nt monomer) and lane 4 (3% self-circularization for the 350 nt monomer). Minor fractions of linear and circular dimeric RNAs are also indicated (lane 2).
    Figure Legend Snippet: Circularization of Mexican axolotl self-cleaved retrozyme RNA. Purified linear monomeric RNA resulting from double self-cleavage of DRtz353 (see Figure 4C ) was run directly in a denaturing gel (lane 1), after 1h incubation with RtcB ligase in its corresponding buffer (lane 2), in RtcB buffer without protein ligase (lane 3), and in 50 mM Mg 2+ buffer (lane 4). Circular RNAs can be readily detected in lane 2 (up to 80% circularization for the 350 nt monomer) and lane 4 (3% self-circularization for the 350 nt monomer). Minor fractions of linear and circular dimeric RNAs are also indicated (lane 2).

    Techniques Used: Purification, Incubation

    Related Articles

    Purification:

    Article Title: Highly efficient expression of circular RNA aptamers in cells using autocatalytic transcripts
    Article Snippet: The products were cleaned by phenol chloroform extraction using heavy phase-lock tubes (Quantabio 2302830). .. 10 pmol of this purified T4-PNK-treated RNA or of the gel purified RNA was ligated using RtcB Ligase (New England Biolabs M0458) for 1 h at 37 °C. ..

    Ligation:

    Article Title: Small circRNAs with self-cleaving ribozymes are highly expressed in diverse metazoan transcriptomes
    Article Snippet: RNA circularization experiments For in vitro analysis of the tRNA ligation and self-ligation capabilities of RNA retrozymes, monomeric retrozyme RNAs that resulted from double self-cleavage after transcription (either in the presence or in the absence of [α-32P]ATP) of dimeric constructs were purified from 5% polyacrylamide gels under denaturing conditions (8 M urea, 1× TBE). .. For the assays of RtcB ligation and self-ligation, 1–10 ng of gel-purified radiolabelled retrozyme RNAs, or 1–4 μg for non-radiolabelled RNAs, were firstly denatured at 95°C for 1 min, cooled down to 25°C (either 0.5°C/s or 1°C/min, with similar results), and then incubated either in the presence of RtcB ligase in its corresponding buffer for 1 h at 37°C (New England Biolabs) or in 50 mM Tris–HCl, pH 8 and 10 to 50 mM MgCl2 for 1 h at 25°C, respectively. ..

    Article Title: Small circRNAs with self-cleaving ribozymes are frequently expressed in metazoan transcriptomes
    Article Snippet: RNA circularization experiments For in vitro analysis of the tRNA ligation and self-ligation capabilities of RNA retrozymes, monomeric retrozyme RNAs that resulted from double self-cleavage after transcription (either in the presence or in the absence of [α-32P]ATP) of dimeric constructs were purified from 5 % polyacrylamide gels under denaturing conditions (8 M urea, 1×TBE). .. For the assays of RtcB ligation, 1 to 10 ng of gel-purified retrozyme RNA were ligated using RtcB ligase in its corresponding buffer for 1 h at 37 °C (New England Biolabs). .. Best results for self-ligation assays were obtained with 1 to 10 ng of gel-purified retrozyme RNA in 50 mM Tris–HCl, pH 8, which were firstly denatured at 95 °C for 1 min, slowly cooled down to 25 °C (1 °C decrease every 2 seconds), and then incubated in the presence of 10 to 50 mM MgCl2 for 1h at 25 °C.

    Article Title: Ligation of 2′, 3′‐cyclic phosphate RNAs for the identification of microRNA binding sites
    Article Snippet: .. RtcB ligation assays One microlitre 10 μm annealed RNA duplexes (0.5 μm final) were mixed with 2 μL 10× RtcB reaction buffer (New England Biolabs), 1 μL 15 μm RtcB (0.75 μm final; New England Biolabs, M0458S), 2 μL 1 mm GTP, 2 μL 10 mm MnCl2 and 12 μL RNase‐free H2O (total volume = 20 μL) and incubated at 37 °C for 1 h. Samples were analyzed by 15% dPAGE and visualized with SYBR Gold Nucleic Acid Gel Stain (Thermo Fisher). .. ResultsIn this study, we aimed to investigate enzymatic ligation as an alternative to cross‐linking in order to capture miRNA‐mRNA interactions, using a new class of miRNA mimics functionalized with 2′, 3′‐cyclic phosphate groups (miRNA > p; Fig. ).

    Incubation:

    Article Title: Small circRNAs with self-cleaving ribozymes are highly expressed in diverse metazoan transcriptomes
    Article Snippet: RNA circularization experiments For in vitro analysis of the tRNA ligation and self-ligation capabilities of RNA retrozymes, monomeric retrozyme RNAs that resulted from double self-cleavage after transcription (either in the presence or in the absence of [α-32P]ATP) of dimeric constructs were purified from 5% polyacrylamide gels under denaturing conditions (8 M urea, 1× TBE). .. For the assays of RtcB ligation and self-ligation, 1–10 ng of gel-purified radiolabelled retrozyme RNAs, or 1–4 μg for non-radiolabelled RNAs, were firstly denatured at 95°C for 1 min, cooled down to 25°C (either 0.5°C/s or 1°C/min, with similar results), and then incubated either in the presence of RtcB ligase in its corresponding buffer for 1 h at 37°C (New England Biolabs) or in 50 mM Tris–HCl, pH 8 and 10 to 50 mM MgCl2 for 1 h at 25°C, respectively. ..

    Article Title: Ligation of 2′, 3′‐cyclic phosphate RNAs for the identification of microRNA binding sites
    Article Snippet: .. RtcB ligation assays One microlitre 10 μm annealed RNA duplexes (0.5 μm final) were mixed with 2 μL 10× RtcB reaction buffer (New England Biolabs), 1 μL 15 μm RtcB (0.75 μm final; New England Biolabs, M0458S), 2 μL 1 mm GTP, 2 μL 10 mm MnCl2 and 12 μL RNase‐free H2O (total volume = 20 μL) and incubated at 37 °C for 1 h. Samples were analyzed by 15% dPAGE and visualized with SYBR Gold Nucleic Acid Gel Stain (Thermo Fisher). .. ResultsIn this study, we aimed to investigate enzymatic ligation as an alternative to cross‐linking in order to capture miRNA‐mRNA interactions, using a new class of miRNA mimics functionalized with 2′, 3′‐cyclic phosphate groups (miRNA > p; Fig. ).

    Staining:

    Article Title: Ligation of 2′, 3′‐cyclic phosphate RNAs for the identification of microRNA binding sites
    Article Snippet: .. RtcB ligation assays One microlitre 10 μm annealed RNA duplexes (0.5 μm final) were mixed with 2 μL 10× RtcB reaction buffer (New England Biolabs), 1 μL 15 μm RtcB (0.75 μm final; New England Biolabs, M0458S), 2 μL 1 mm GTP, 2 μL 10 mm MnCl2 and 12 μL RNase‐free H2O (total volume = 20 μL) and incubated at 37 °C for 1 h. Samples were analyzed by 15% dPAGE and visualized with SYBR Gold Nucleic Acid Gel Stain (Thermo Fisher). .. ResultsIn this study, we aimed to investigate enzymatic ligation as an alternative to cross‐linking in order to capture miRNA‐mRNA interactions, using a new class of miRNA mimics functionalized with 2′, 3′‐cyclic phosphate groups (miRNA > p; Fig. ).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    New England Biolabs rtcb ligase
    Circularization of Mexican axolotl self-cleaved <t>retrozyme</t> RNA. Purified linear monomeric RNA resulting from double self-cleavage of DRtz353 (see Figure 4C ) was run directly in a denaturing gel (lane 1), after 1h incubation with <t>RtcB</t> ligase in its corresponding buffer (lane 2), in RtcB buffer without protein ligase (lane 3), and in 50 mM Mg 2+ buffer (lane 4). Circular RNAs can be readily detected in lane 2 (up to 80% circularization for the 350 nt monomer) and lane 4 (3% self-circularization for the 350 nt monomer). Minor fractions of linear and circular dimeric RNAs are also indicated (lane 2).
    Rtcb Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rtcb ligase/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rtcb ligase - by Bioz Stars, 2021-07
    94/100 stars
      Buy from Supplier

    Image Search Results


    Circularization of Mexican axolotl self-cleaved retrozyme RNA. Purified linear monomeric RNA resulting from double self-cleavage of DRtz353 (see Figure 4C ) was run directly in a denaturing gel (lane 1), after 1h incubation with RtcB ligase in its corresponding buffer (lane 2), in RtcB buffer without protein ligase (lane 3), and in 50 mM Mg 2+ buffer (lane 4). Circular RNAs can be readily detected in lane 2 (up to 80% circularization for the 350 nt monomer) and lane 4 (3% self-circularization for the 350 nt monomer). Minor fractions of linear and circular dimeric RNAs are also indicated (lane 2).

    Journal: bioRxiv

    Article Title: Small circRNAs with self-cleaving ribozymes are frequently expressed in metazoan transcriptomes

    doi: 10.1101/721605

    Figure Lengend Snippet: Circularization of Mexican axolotl self-cleaved retrozyme RNA. Purified linear monomeric RNA resulting from double self-cleavage of DRtz353 (see Figure 4C ) was run directly in a denaturing gel (lane 1), after 1h incubation with RtcB ligase in its corresponding buffer (lane 2), in RtcB buffer without protein ligase (lane 3), and in 50 mM Mg 2+ buffer (lane 4). Circular RNAs can be readily detected in lane 2 (up to 80% circularization for the 350 nt monomer) and lane 4 (3% self-circularization for the 350 nt monomer). Minor fractions of linear and circular dimeric RNAs are also indicated (lane 2).

    Article Snippet: For the assays of RtcB ligation, 1 to 10 ng of gel-purified retrozyme RNA were ligated using RtcB ligase in its corresponding buffer for 1 h at 37 °C (New England Biolabs).

    Techniques: Purification, Incubation