t4 dna ligase (Thermo Fisher)


Name:
T4 DNA Ligase
Description:
Thermo Scientific T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5 phosphate and 3 hydroxyl termini in duplex DNA or RNA The enzyme repairs single strand nicks in duplex DNA RNA or DNA RNA hybrids It also joins DNA fragments with either cohesive or blunt termini but has no activity on single stranded nucleic acids T4 DNA Ligase requires ATP as a cofactor Highlights• Active in Themo Scientific restriction enzyme PCR and RT buffers when supplemented with ATP • Fast sticky end ligation is completed in 10 minutes at room temperature• Supplied with PEG solution for efficient blunt end ligationApplications• Cloning of restriction enzyme generated DNA fragments• Cloning of PCR products• Joining of double stranded oligonucleotide linkers or adaptors to DNA• Site directed mutagenesis• Amplified fragment length polymorphism AFLP • Ligase mediated RNA detection see Reference 3 • Nick repair in duplex DNA RNA or DNA RNA hybrids• Self circularization of linear DNA Includes• T4 DNA Ligase• 10X T4 DNA Ligase Buffer• 50 PEG SolutionNotes• Binding of T4 DNA Ligase to DNA may result in a band shift in agarose gels To avoid this incubate samples with 6X DNA Loading Dye SDS Solution at 70°C for 5 min or 65°C for 10 minutes and chill on ice prior to electrophoresis • The volume of the ligation reaction mixture should not exceed 10 of the competent cell volume in the transformation process • Prior to electro transformation remove T4 DNA Ligase from the ligation mixture using spin columns or chloroform extraction The extracted DNA can be further precipitated with ethanol
Catalog Number:
el0013
Price:
None
Category:
Proteins Enzymes Peptides
Applications:
Cloning|Restriction Enzyme Cloning
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Structured Review
Thermo Scientific T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5 phosphate and 3 hydroxyl termini in duplex DNA or RNA The enzyme repairs single strand nicks in duplex DNA RNA or DNA RNA hybrids It also joins DNA fragments with either cohesive or blunt termini but has no activity on single stranded nucleic acids T4 DNA Ligase requires ATP as a cofactor Highlights• Active in Themo Scientific restriction enzyme PCR and RT buffers when supplemented with ATP • Fast sticky end ligation is completed in 10 minutes at room temperature• Supplied with PEG solution for efficient blunt end ligationApplications• Cloning of restriction enzyme generated DNA fragments• Cloning of PCR products• Joining of double stranded oligonucleotide linkers or adaptors to DNA• Site directed mutagenesis• Amplified fragment length polymorphism AFLP • Ligase mediated RNA detection see Reference 3 • Nick repair in duplex DNA RNA or DNA RNA hybrids• Self circularization of linear DNA Includes• T4 DNA Ligase• 10X T4 DNA Ligase Buffer• 50 PEG SolutionNotes• Binding of T4 DNA Ligase to DNA may result in a band shift in agarose gels To avoid this incubate samples with 6X DNA Loading Dye SDS Solution at 70°C for 5 min or 65°C for 10 minutes and chill on ice prior to electrophoresis • The volume of the ligation reaction mixture should not exceed 10 of the competent cell volume in the transformation process • Prior to electro transformation remove T4 DNA Ligase from the ligation mixture using spin columns or chloroform extraction The extracted DNA can be further precipitated with ethanol
https://www.bioz.com/result/t4 dna ligase/product/Thermo Fisher
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Incubation:Article Title: Autophosphorylation-dependent remodeling of the DNA-dependent protein kinase catalytic subunit regulates ligation of DNA ends Article Snippet: .. Reactions were started by the addition of 0.5 U of Purification:Article Title: FSH1 regulates the phenotype and pathogenicity of the pathogenic dermatophyte Microsporum canis Article Snippet: To construct the final FSH1 double-stranded RNA interference (dsRNAi) plasmid pCB309-PFUFT, three steps of ligation were performed as follows. .. First, the purified product of PCR for the FSH1 gene was ligated into pUC-PUT following DNA digestion with Xho I and Hin dIII, and ligated by Polymerase Chain Reaction:Article Title: FSH1 regulates the phenotype and pathogenicity of the pathogenic dermatophyte Microsporum canis Article Snippet: To construct the final FSH1 double-stranded RNA interference (dsRNAi) plasmid pCB309-PFUFT, three steps of ligation were performed as follows. .. First, the purified product of PCR for the FSH1 gene was ligated into pUC-PUT following DNA digestion with Xho I and Hin dIII, and ligated by Modification:Article Title: In vitro repair of complex unligatable oxidatively induced DNA double-strand breaks by human cell extracts Article Snippet: Finally, due to its relatively speedy processing times and lack of radioactive components, this assay can be easily scaled up to levels appropriate for biochemical analysis and protein purification or for high throughput screening methods. .. Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum, non-essential amino acids (10 mM), glutamine (100 mM), penicillin/streptomycin (10 000 U/ml), Saccharomyces cerevisiae tRNA and Ligation:Article Title: Practical Synthesis of Cap‐4 RNA Article Snippet: Enzymatic ligation of cap‐4 RNA . .. The 38‐nt T. cruzi cap‐4 RNA was prepared by splinted enzymatic ligation of an 11‐nt cap‐4 RNA and a chemically synthesized 5′‐phosphorylated 27‐nt RNA by using Article Title: Profiling non-lysyl tRNAs in HIV-1 Article Snippet: The ligation reaction relies on an 8-bp RNA:DNA hybrid helix containing a Cy3 or Cy5 fluorophore pre-attached to the loop and an overhang complementary to the universally conserved 3′CCA nucleotides present in all tRNAs ( ). .. The ligation reaction was carried out overnight (∼16 h) at 16°C with 0.13 μg/μL total RNA in 1X T4 DNA ligase buffer, 0.5 U/μL Synthesized:Article Title: Practical Synthesis of Cap‐4 RNA Article Snippet: Enzymatic ligation of cap‐4 RNA . .. The 38‐nt T. cruzi cap‐4 RNA was prepared by splinted enzymatic ligation of an 11‐nt cap‐4 RNA and a chemically synthesized 5′‐phosphorylated 27‐nt RNA by using Labeling:Article Title: Profiling non-lysyl tRNAs in HIV-1 Article Snippet: The ligation reaction relies on an 8-bp RNA:DNA hybrid helix containing a Cy3 or Cy5 fluorophore pre-attached to the loop and an overhang complementary to the universally conserved 3′CCA nucleotides present in all tRNAs ( ). .. The ligation reaction was carried out overnight (∼16 h) at 16°C with 0.13 μg/μL total RNA in 1X T4 DNA ligase buffer, 0.5 U/μL Mass Spectrometry:Article Title: Detection of Ligation Products of DNA Linkers with 5?-OH Ends by Denaturing PAGE Silver Stain Article Snippet: When these manufacturers were questioned, they stated that their T4 DNA ligases had very high quality and it was very unlikely that there would be PNK in their ligases because their ligases were produced by using E. coli cells and production lines that were different from those for T4 PNK. .. A quality inspection report of |