T4 RNA Ligase 1 (ssRNA Ligase, 10 U/μL) _ 14651ES

T4 RNA Ligase 1, also known as T4 RNA ligase, is an ATP-dependent enzyme that catalyzes the formation of a phosphodiester bond between the 5'-P terminus and the 3'-OH terminus of single-stranded RNA (ssRNA), single-stranded DNA (ssDNA), or mononucleotide molecules. T4 RNA Ligase 1 can perform ligation between ssRNA molecules (most efficiently), between ssDNA and ssRNA (with lower efficiency), and can also be used for ligation between ssDNA molecules (with very low efficiency). Applications ssRNA/ssDNA Ligation 5' adapters in miRNA library construction for NGS Circular RNA Preparation Specifications Cat.No. 14651ES80/ 14651ES90 Size 1,000 U / 5,000 U Unit Definition One unit is defined as the amount of enzyme required to convert 1 nanomole of 5 -[32P] rA16 into a phosphatase-resistant form in 30 minutes at 37°C. Source Recombinant protein expressed by E. coli is derived from the T4 bacteriophage. Reaction conditions 10×T4 RL1 Reaction Buffer: 500 mM Tris-HCl (pH 7.5 at 25℃), 100 mM MgCl2, 20 mM DTT; Optimal incubation at 37°C. Inactivation conditions Heating at 65°C for 15 minutes or boiling for 2 minutes can inactivate T4 RNA Ligase 1. Alternatively, adding a final concentration of 50 mM EDTA pH 8.0 can terminate the reaction of T4 RNA Ligase 1. Components Components No. Name 14651ES80 14651ES90 14651-A T4 RNA Ligase 1 (10 U/μL) 100 μL 500 μL 14651-B 10×T4 RL1 Reaction Buffer 500 μL 2 ×1.5 mL Figures • High Ligation efficiency Yeasen T4 RNA Ligase 1 and T4 RNA Ligase 1 from another brand (Supplier A*) were added at different amounts to ligate a 21-nt ssDNA substrate (1 μM final concentration) and a 26-nt ssRNA substrate (0.5 μM final concentration). Yeasen T4 RNA Ligase 1 efficiently ligated both the ssRNA and ssDNA substrates, with better ligation performance than Supplier A*. Figure 1. Ligation efficiency of T4 RNA Ligase 1 • Significantly lower nicking endonuclease contamination Figure 2. Nicking endonuclease contamination assay Note: 30 U of T4 RNA Ligase 1 from Yeasen and Brand N* was incubated with plasmid DNA, and changes in DNA bands were compared by agarose gel electrophoresis. All three lots of Yeasen T4 RNA Ligase 1 showed significantly lower nicking endonuclease contamination than Brand N*. • No exonuclease contamination Figure 3. Exonuclease contamination assay Note: 30 U of T4 RNA Ligase 1 was incubated with substrate DNA, and changes in DNA bands were compared by agarose gel electrophoresis. No exonuclease contamination was detected in any of the three lots of Yeasen T4 RNA Ligase 1. • No RNase contamination Figure 4. RNase contamination assay Note: 10 U of T4 RNA Ligase 1 was incubated with substrate RNA, and changes in RNA bands were compared by agarose gel electrophoresis. No RNase contamination was detected in any of the three lots of Yeasen T4 RNA Ligase 1.