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Thermo Scientific™ T4 DNA Ligase PROMO
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA.
Marca: Thermo Scientific™ EL0016-RMD123
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Descripción
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 ligation
Applications
- 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.
Notes
- 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 by spin column or chloroform extraction. The extracted DNA can be further precipitated with ethanol.
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