T4 DNA Ligase, HC (30 U/µL)/T4 DNA连接酶，HC

货号：EL0013

规格：5000 units

价格：3375

产品类型：酶及抑制剂

品牌：Thermo Fisher

描述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 requiresATPas 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 (seeReference 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 with6X DNA Loading Dye & SDS Solutionat 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.

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