T4 ligation buffer. 8 at 25°C), 100mM MgCl 2, 100mM DTT and 10mM ATP. The 10X buffer has been tested for absence of endo-, exodeoxyribonucleases, ribonucleases and functionally tested in DNA ligation. A 1X concentration assures optimal enzyme activity. 10X reaction buffer supplied with T4 DNA Ligase. Oct 11, 2023 ยท The DNA ligation buffer for T4 DNA ligase contains 10mM part of ATP which provides two ATP energy molecules in each reaction. * The T4 DNA Ligase Buffer should be thawed and resuspended at room temperature. The enzyme will not join single-stranded nucleic acids. The 10x T4 DNA Ligase Buffer does not contain PEG and is compatible with standard ligation protocols that do not specify the use of a rapid, fast or quick format buffer. Use NEBioCalculator to calculate molar ratios. . The Thermo Scientific 10X T4 DNA Ligase buffer is used with T4 DNA Ligase. T4 DNA Ligase catalyzes the formation of phosphodiester bonds in the presence of ATP between double-stranded DNAs with 3´ hydroxyl and 5´ phosphate termini. The unique T4 DNA Ligase buffer optimizes ligation, which can be performed in 5 minutes (1). T4 DNA Ligase is provided with 10X Reaction Buffer: 300mM Tris-HCl (pH 7. Gently mix the reaction by pipetting up and down and microfuge briefly. T4 DNA Ligase catalyzes the joining of two cohesive- or blunt-ended strands of DNA between the 5´-phosphate and the 3´-hydroxyl groups of adjacent nucleotides. The activity of ATP degrades upon repeated freeze/thawing. For cohesive (sticky) ends, incubate at 16°C overnight or room temperature for 10 minutes. ltool lrzs acdyvqo upvbh ziw paktyb hrgvhvfq xbps eqij fvhfcs