59,00 € – 180,00 €ATP-dependent recombinant enzyme used for molecular cloning, site-directed mutagenesis, nick repair in duplex DNA, RNA or DNA/RNA hybrids, Ligation Mediated PCR; concentration 5 U/μl
- Additional information
- Product Information
- Protocol & Manual
- Technical Tips
- Quality & Safety
500 U, 2500U
T4 DNA Ligase is an ATP-dependent recombinant enzyme isolated from Escherichia coli strain used to clone DNA. T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5′-phosphate and 3′-hydroxyl termini in duplex DNA or RNA. It will join both blunt-ended and cohesive-ended restriction fragments of DNA, as well as repair single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
- ATP-dependent recombinant DNA ligases over-expressed in E. coli.
- Available also in special formulation for very fast and efficient ligation of DNA fragments with compatible cohesive or blunt ends in 5 and 15 minutes respectively.
- Molecular cloning of PCR products or restriction fragments.
- Joining double-stranded oligonucleotide linkers or adaptors to DNA.
- Site-directed mutagenesis.
- Nick repair in duplex DNA, RNA or DNA/RNA hybrids.
- Self-circularization of linear DNA.
- LM PCR methods (Ligation Mediated PCR).
One (Weiss) unit of T4 DNA Ligase catalyzes the conversion of 1 nmol of 32P from pyrophosphate into Norit-adsorbable material in 30 minutes at 37°C.
One Weiss unit is equivalent an approximately 200 cohesive end units.
Protocol & Manual
T4 DNA Ligase (EN11) - Manual
- The 10x Ligation Buffer and ATP Solution should be thawed and resuspended at room temperature.
- For blunt-end ligations, use higher quantities of both the vector and the insert DNA.
- For sticky (cohesive)-end ligations, we recommend heating both the vector and the insert DNA prior to ligation.
- The electrotransformation efficiency may be improved by heat inactivation of the T4 DNA Ligase and purifi cation of the DNA by means of a spin column purification method (EXTRACTME DNA CLEAN-UP kit or
EXTRACTME DNA GEL-OUT kit).
- We recommend using a 3 – 10 molar excess of insert DNA over vector DNA. To calculate the optimal amounts of insert DNA in a ligation reaction,use the following equation:
Example: If using 20 ng of a vector plasmid (4 kb), for a 5:1 molar ratio of insert : vector, you will require the following quantity of 1 kb insert:
- The enzyme is inhibited by >200 mM NaCl or KCl concentrations.
- Inactivate enzyme at 65°C for 10 minutes or at 70°C for 5 minutes.
Quality & Safety
The product was tested in ligation of HindIII-cut lambda DNA with a different amounts of enzyme. The product is free of unspecific DNA nucleases.
T4 DNA Ligase (EN11) - MSDS
T4 DNA Ligase (EN11) - CoA 1.2018 L.686771
Sensitive and inexpensive digital DNA analysis by microfluidic enrichment of rolling circle amplified single-molecules
Authors: Malte Kühnemund, Iván Hernández-Neuta, Mohd Istiaq Sharif, Matteo Cornaglia, Martin A.M. Gijs and Mats Nilsson Products: T4 DNA ligase (EN11), BSA (Bovine Serum Albumin) (EN17), ATP Year: 2017 Source: Nucleic Acids Research, 2017, Vol. 45, No. 8 e59
Targeted DNA sequencing and in situ mutation analysis using mobile phone microscopy
Authors: Malte Kühnemund Qingshan Wei, Evangelia Darai, Yingjie Wang, Iván Hernández-Neuta, Zhao Yang, Derek Tseng, Annika Ahlford, Lucy Mathot, Tobias Sjöblom, Aydogan Ozcan & Mats Nilsson Products: T4 DNA Ligase (EN11), TRANSCRIPTME RNA Kit (RT31), RNase H (RT34), dNTPs Year: 2017 Source: NATURE COMMUNICATIONS
Next-generation bis-locked nucleic acids with stacking linker and 2-glycylamino-LNA show enhanced DNA invasion into supercoiled duplexes
Authors: Sylvain Geny, Pedro M. D. Moreno, Tomasz Krzywkowski, Olof Gissberg, Nicolai K. Andersen, Abdirisaq J. Isse, Amro M. El-Madani, Chenguang Lou, Y. Vladimir Pabon, Brooke A. Anderson, Eman M. Zaghloul, Rula Zain, Patrick J. Hrdlicka, Per T. Jørgensen, Mats Nilsson, Karin E. Lundin, Erik B. Pedersen, Jesper Wengel and C. I. Edvard Smith Products: T4 DNA ligase (EN11), ATP Year: 2016 Source: Nucleic Acids Research, 2016, Vol. 44, No. 5 2007–2019