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TaKaRa t4 rna ligase
T4 Rna Ligase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 95/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/t4 rna ligase/product/TaKaRa
Average 95 stars, based on 74 article reviews
Price from $9.99 to $1999.99
t4 rna ligase - by Bioz Stars, 2020-04
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Selection:

Article Title: MicroRNAs and their targeted genes associated with phase changes of stem explants during tissue culture of tea plant
Article Snippet: In brief, sRNAs were firstly ligated with 3′ and 5′RNA adapters by using T4 RNA Ligase 1 & 2 (truncated) (Takara, Dalian, China). .. Size selection of the PCR products were performed on PAGE gel, and sRNA libraries were recovered by gel-cutting and purified through AMPure XP Kit (Beckman Coulter, Australia).

Construct:

Article Title: MicroRNAs and their targeted genes associated with phase changes of stem explants during tissue culture of tea plant
Article Snippet: Small RNA library preparation and sequencing After qualification of the RNAs, 1.5 μg RNAs of each sample were used to construct the sequencing library with a NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB, USA) according to the manufacturer’s recommendations. .. In brief, sRNAs were firstly ligated with 3′ and 5′RNA adapters by using T4 RNA Ligase 1 & 2 (truncated) (Takara, Dalian, China).

Purification:

Article Title: MicroRNAs and their targeted genes associated with phase changes of stem explants during tissue culture of tea plant
Article Snippet: In brief, sRNAs were firstly ligated with 3′ and 5′RNA adapters by using T4 RNA Ligase 1 & 2 (truncated) (Takara, Dalian, China). .. Size selection of the PCR products were performed on PAGE gel, and sRNA libraries were recovered by gel-cutting and purified through AMPure XP Kit (Beckman Coulter, Australia).

Sequencing:

Article Title: MicroRNAs and their targeted genes associated with phase changes of stem explants during tissue culture of tea plant
Article Snippet: Paragraph title: Small RNA library preparation and sequencing ... In brief, sRNAs were firstly ligated with 3′ and 5′RNA adapters by using T4 RNA Ligase 1 & 2 (truncated) (Takara, Dalian, China).

Polyacrylamide Gel Electrophoresis:

Article Title: MicroRNAs and their targeted genes associated with phase changes of stem explants during tissue culture of tea plant
Article Snippet: In brief, sRNAs were firstly ligated with 3′ and 5′RNA adapters by using T4 RNA Ligase 1 & 2 (truncated) (Takara, Dalian, China). .. Size selection of the PCR products were performed on PAGE gel, and sRNA libraries were recovered by gel-cutting and purified through AMPure XP Kit (Beckman Coulter, Australia).

Polymerase Chain Reaction:

Article Title: MicroRNAs and their targeted genes associated with phase changes of stem explants during tissue culture of tea plant
Article Snippet: In brief, sRNAs were firstly ligated with 3′ and 5′RNA adapters by using T4 RNA Ligase 1 & 2 (truncated) (Takara, Dalian, China). .. PCR amplifications were performed with primers that annealed to the ends of the adapters.

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    TaKaRa t4 rna ligase
    ( A ) Schematic illustration of the formation of streptavidin–tRNA fusion using puromycin–tRNA, which contains a puromycin moiety in the place of 3′ terminal aminoacyl-adenosine and a four-base anticodon CCCG. The puromycin–tRNA binds to ribosomal A site and accepts a streptavidin polypeptide chain as an analog of aminoacyl-tRNA in response to a four-base CGGG codon at 3′ terminus of streptavidin mRNA in a cell-free translation. The resulting streptavidin–puromycin–tRNA may be translocated to the P-site. In this case, the next aminoacyl-tRNA binds to the vacant ribosomal A site, but can not accept the polypeptide chain because of the amide bond of puromycin–tRNA. The resulting streptavidin–tRNA fusion is released from the ribosome complex by the addition of EDTA. ( B ) Schematic illustration of the in vitro selection system of tRNAs. Step 1, a DNA pool encoding tRNAs containing a four-base anticodon CCCG is transcribed by T7 RNA polymerase to tRNA(-CA) pool. Step 2, the tRNA(-CA) pool is ligated with pdCp-Puromycin by <t>T4</t> RNA ligase to generate puromycin–tRNA. Step 3, a streptavidin mRNA containing a four-base CGGG codon at C-terminus is translated in an E.coli cell-free translation system in the presence of the puromycin–tRNA. Puromycin–tRNAs that successfully decode the CGGG codon form ribosome–mRNA–streptavidin–tRNA complex. Step 4, the streptavidin–tRNA fusion is dissociated from the complex by the addition of EDTA. Step 5, the streptavidin–tRNA fusion is recovered with biotin-coated magnetic beads. Step 6, the streptavidin–tRNA fusion is dissociated from the beads, and then the tRNA moiety is subjected to RT–PCR. Step 7, the tRNA genes are regenerated by overlap-extension PCR with a T7 promoter primer, which are used as template DNAs in the next round of selection.
    T4 Rna Ligase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 rna ligase/product/TaKaRa
    Average 99 stars, based on 74 article reviews
    Price from $9.99 to $1999.99
    t4 rna ligase - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

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    ( A ) Schematic illustration of the formation of streptavidin–tRNA fusion using puromycin–tRNA, which contains a puromycin moiety in the place of 3′ terminal aminoacyl-adenosine and a four-base anticodon CCCG. The puromycin–tRNA binds to ribosomal A site and accepts a streptavidin polypeptide chain as an analog of aminoacyl-tRNA in response to a four-base CGGG codon at 3′ terminus of streptavidin mRNA in a cell-free translation. The resulting streptavidin–puromycin–tRNA may be translocated to the P-site. In this case, the next aminoacyl-tRNA binds to the vacant ribosomal A site, but can not accept the polypeptide chain because of the amide bond of puromycin–tRNA. The resulting streptavidin–tRNA fusion is released from the ribosome complex by the addition of EDTA. ( B ) Schematic illustration of the in vitro selection system of tRNAs. Step 1, a DNA pool encoding tRNAs containing a four-base anticodon CCCG is transcribed by T7 RNA polymerase to tRNA(-CA) pool. Step 2, the tRNA(-CA) pool is ligated with pdCp-Puromycin by T4 RNA ligase to generate puromycin–tRNA. Step 3, a streptavidin mRNA containing a four-base CGGG codon at C-terminus is translated in an E.coli cell-free translation system in the presence of the puromycin–tRNA. Puromycin–tRNAs that successfully decode the CGGG codon form ribosome–mRNA–streptavidin–tRNA complex. Step 4, the streptavidin–tRNA fusion is dissociated from the complex by the addition of EDTA. Step 5, the streptavidin–tRNA fusion is recovered with biotin-coated magnetic beads. Step 6, the streptavidin–tRNA fusion is dissociated from the beads, and then the tRNA moiety is subjected to RT–PCR. Step 7, the tRNA genes are regenerated by overlap-extension PCR with a T7 promoter primer, which are used as template DNAs in the next round of selection.

    Journal: Nucleic Acids Research

    Article Title: In vitro selection of tRNAs for efficient four-base decoding to incorporate non-natural amino acids into proteins in an Escherichia coli cell-free translation system

    doi: 10.1093/nar/gkl087

    Figure Lengend Snippet: ( A ) Schematic illustration of the formation of streptavidin–tRNA fusion using puromycin–tRNA, which contains a puromycin moiety in the place of 3′ terminal aminoacyl-adenosine and a four-base anticodon CCCG. The puromycin–tRNA binds to ribosomal A site and accepts a streptavidin polypeptide chain as an analog of aminoacyl-tRNA in response to a four-base CGGG codon at 3′ terminus of streptavidin mRNA in a cell-free translation. The resulting streptavidin–puromycin–tRNA may be translocated to the P-site. In this case, the next aminoacyl-tRNA binds to the vacant ribosomal A site, but can not accept the polypeptide chain because of the amide bond of puromycin–tRNA. The resulting streptavidin–tRNA fusion is released from the ribosome complex by the addition of EDTA. ( B ) Schematic illustration of the in vitro selection system of tRNAs. Step 1, a DNA pool encoding tRNAs containing a four-base anticodon CCCG is transcribed by T7 RNA polymerase to tRNA(-CA) pool. Step 2, the tRNA(-CA) pool is ligated with pdCp-Puromycin by T4 RNA ligase to generate puromycin–tRNA. Step 3, a streptavidin mRNA containing a four-base CGGG codon at C-terminus is translated in an E.coli cell-free translation system in the presence of the puromycin–tRNA. Puromycin–tRNAs that successfully decode the CGGG codon form ribosome–mRNA–streptavidin–tRNA complex. Step 4, the streptavidin–tRNA fusion is dissociated from the complex by the addition of EDTA. Step 5, the streptavidin–tRNA fusion is recovered with biotin-coated magnetic beads. Step 6, the streptavidin–tRNA fusion is dissociated from the beads, and then the tRNA moiety is subjected to RT–PCR. Step 7, the tRNA genes are regenerated by overlap-extension PCR with a T7 promoter primer, which are used as template DNAs in the next round of selection.

    Article Snippet: T4 RNA ligase, Bca BEST RNA PCR kit ver1.1, GelStar Nucleic Acid Stain and ribonuclease inhibitor were from TaKaRa BIO.

    Techniques: In Vitro, Selection, Magnetic Beads, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction

    Gel-electrophoresis of ligation products. The ligation products were electrophoresed on 8M Urea 10% PAGE at 65 ºC and were visualized with fluorescence of FITC and then visualized after staining by SYBR Green II using an imager. Lane CY: control, Linker-Y. Lane CS: control, mRNA without ligation. Lane CR: control, Linker-S. Lane Y: Y-ligation with T4 RNA ligase. Lane R: splint ligation with T4 RNA ligase. Lane D: splint ligation with T4 DNA ligase.

    Journal: Biological Procedures Online

    Article Title: An Efficient Ligation Method in the Making of an in vitro Virus for in vitro Protein Evolution

    doi: 10.1251/bpo33

    Figure Lengend Snippet: Gel-electrophoresis of ligation products. The ligation products were electrophoresed on 8M Urea 10% PAGE at 65 ºC and were visualized with fluorescence of FITC and then visualized after staining by SYBR Green II using an imager. Lane CY: control, Linker-Y. Lane CS: control, mRNA without ligation. Lane CR: control, Linker-S. Lane Y: Y-ligation with T4 RNA ligase. Lane R: splint ligation with T4 RNA ligase. Lane D: splint ligation with T4 DNA ligase.

    Article Snippet: Ligation reaction between mRNA and puromycin-linker We compared three puromycin-linker attachment methods, which are the splint ligation with T4 DNA ligase , the splint ligation with T4 RNA ligase and the Y-ligation with T4 RNA ligase (Fig. ).

    Techniques: Nucleic Acid Electrophoresis, Ligation, Polyacrylamide Gel Electrophoresis, Fluorescence, Staining, SYBR Green Assay

    Time course of ligation reactions. Concentrations were [mRNA] = 0.5 µM and [T4 RNA ligase] = 1 U/µl in the enzyme-supplier's buffer with 10% DMSO. At 25 ºC.(a) Y-ligation, [mRNA]:[Linker-Y] = 1:1.5(b) Splint ligation with T4 RNA ligase as compared with Y-ligation, [mRNA]:[Splint]:[Linker-S] = 1:1:1.5

    Journal: Biological Procedures Online

    Article Title: An Efficient Ligation Method in the Making of an in vitro Virus for in vitro Protein Evolution

    doi: 10.1251/bpo33

    Figure Lengend Snippet: Time course of ligation reactions. Concentrations were [mRNA] = 0.5 µM and [T4 RNA ligase] = 1 U/µl in the enzyme-supplier's buffer with 10% DMSO. At 25 ºC.(a) Y-ligation, [mRNA]:[Linker-Y] = 1:1.5(b) Splint ligation with T4 RNA ligase as compared with Y-ligation, [mRNA]:[Splint]:[Linker-S] = 1:1:1.5

    Article Snippet: Ligation reaction between mRNA and puromycin-linker We compared three puromycin-linker attachment methods, which are the splint ligation with T4 DNA ligase , the splint ligation with T4 RNA ligase and the Y-ligation with T4 RNA ligase (Fig. ).

    Techniques: Ligation

    Gel electrophoresis pattern of mRNA-linker ligation. The ligation products reacted with or without prRT- DNA oligomer used as a blocker of the 3'-end of mRNA were electrophoresis on 8 M urea 8 % PAGE at 65 °C and were visualized with fluorescence of (A) SYBR Green II and (B) FITC. Lane M: DNA ladder, Lane Y: ligation product, Lane L-: negative control, reaction product without DNA-linker, Lane E-: negative control, reaction product without T4 RNA ligase. Mobility of the mRNA-linker and the self-ligation product of mRNA are shown to be equivalent.

    Journal: International Journal of Biological Sciences

    Article Title: An mRNA-protein Fusion at N-terminus for Evolutionary Protein Engineering

    doi:

    Figure Lengend Snippet: Gel electrophoresis pattern of mRNA-linker ligation. The ligation products reacted with or without prRT- DNA oligomer used as a blocker of the 3'-end of mRNA were electrophoresis on 8 M urea 8 % PAGE at 65 °C and were visualized with fluorescence of (A) SYBR Green II and (B) FITC. Lane M: DNA ladder, Lane Y: ligation product, Lane L-: negative control, reaction product without DNA-linker, Lane E-: negative control, reaction product without T4 RNA ligase. Mobility of the mRNA-linker and the self-ligation product of mRNA are shown to be equivalent.

    Article Snippet: Ligation between the mRNA and the hydrazide-linker The mRNA (2 µM) was hybridized to the DNA moiety of the hydrazide-linker (4 µM) and prRT- (4 µM) by heating at 95 °C and cooling to 25 °C in 50 µl of T4 RNA ligase buffer (Takara Bio) and ligation reaction was started by adding T4 RNA ligase (40 U, Takara Bio) and ribonuclease inhibitor (40 U, Takara Bio).

    Techniques: Nucleic Acid Electrophoresis, Ligation, Electrophoresis, Polyacrylamide Gel Electrophoresis, Fluorescence, SYBR Green Assay, Negative Control

    Screening cycle of the mRNA-protein fusion in this study. The dsDNA library is transcribed to mRNA. The mRNA is hybridized to the DNA moiety of the linker having hydrazide group and ligated with T4 RNA ligase. Hydrazide group of the ligated product and acetyl group of the phenylalanine derivative that is acylated to sup tRNA are ligated chemically and the modified mRNA is translated. The modified aminoacyl sup tRNA tends to occupy the A-site of ribosome at UAG codon inserted near downstream of initiation codon and the phenylalanine is incorporated into the growing peptide. Thus, linkage between N-terminus of the nascent peptide and 5'-terminus of its mRNA is achieved. Screening of mRNA-peptide fusion library according to property of the displayed peptide and amplify the genotype molecules of the screened fusions by RT-PCR.

    Journal: International Journal of Biological Sciences

    Article Title: An mRNA-protein Fusion at N-terminus for Evolutionary Protein Engineering

    doi:

    Figure Lengend Snippet: Screening cycle of the mRNA-protein fusion in this study. The dsDNA library is transcribed to mRNA. The mRNA is hybridized to the DNA moiety of the linker having hydrazide group and ligated with T4 RNA ligase. Hydrazide group of the ligated product and acetyl group of the phenylalanine derivative that is acylated to sup tRNA are ligated chemically and the modified mRNA is translated. The modified aminoacyl sup tRNA tends to occupy the A-site of ribosome at UAG codon inserted near downstream of initiation codon and the phenylalanine is incorporated into the growing peptide. Thus, linkage between N-terminus of the nascent peptide and 5'-terminus of its mRNA is achieved. Screening of mRNA-peptide fusion library according to property of the displayed peptide and amplify the genotype molecules of the screened fusions by RT-PCR.

    Article Snippet: Ligation between the mRNA and the hydrazide-linker The mRNA (2 µM) was hybridized to the DNA moiety of the hydrazide-linker (4 µM) and prRT- (4 µM) by heating at 95 °C and cooling to 25 °C in 50 µl of T4 RNA ligase buffer (Takara Bio) and ligation reaction was started by adding T4 RNA ligase (40 U, Takara Bio) and ribonuclease inhibitor (40 U, Takara Bio).

    Techniques: Modification, Reverse Transcription Polymerase Chain Reaction