Keterangan
T4 RNA Ligase 2, truncated KQ is a double-point mutant (R55K, K227Q) of the truncated form of T4 RNA Ligase 2. While maintaining high ligation efficiency, it significantly reduces non-specific RNA ligation (e.g., RNA concatemerization or circularization). This enzyme specifically ligates a 5′-pre-adenylated DNA or RNA adapter to the 3′-OH end of an RNA molecule, making it highly suitable for adapter ligation to small RNAs in cDNA library construction.ATP is not required for this reaction, but the adapter must be pre-adenylated at the 5′ end.
Features
- Protein purity ≥ 95%
- Free of detectable nuclease, nicking enzyme, RNase, and phosphatase contamination
- High ligation activity
Application
Small RNA Library preparation
Specifications
|
Cat NO. |
14653ES90 / 14653ES94 / 14653ES98 |
|
Size |
5 KU / 20 KU / 100 KU |
|
Unit Definition |
One unit is defined as the amount of enzyme that ligates 80% of a 31-mer RNA to a pre-adenylated 17-mer DNA in a 20 μL reaction at 25°C for 1 hour. Thus, 200 units = 20 μL reaction under these conditions. |
|
Source |
Recombinant expression in E. coli of a truncated mutant of bacteriophage T4 gene Y10A. |
|
Purity |
≥95% |
|
Reaction Conditions |
Incubate at 25°C for 1 hour. |
|
Heat Inactivation |
Incubate at 65°C for 20 minutes. |
Components
|
Components No. |
Name |
14653ES90 |
14653ES94 |
|
14653-A |
T4 RNA Ligase 2, truncated KQ (200 U/μL) |
25 μL |
100 μL |
|
14653-B |
10× T4 RL Reaction Buffer |
0.5 mL |
1 mL |
Storage
This product should be stored at -25~-15℃ for 2 years.
Figures
1. High ligation activity
Figure 1. Ligation Performance of T4 RNA Ligase 2, Truncated KQ
T4 RNA Ligase 2, Truncated KQ from Yeasen and Supplier A* was tested at varying enzyme inputs in ligation reactions containing a 31-nt single-stranded RNA (ssRNA) substrate (Substrate 1) and a 17-nt pre-adenylated ssRNA substrate (Substrate 2). Results demonstrate that Yeasen’s enzyme efficiently ligates ssRNA substrates, delivering performance comparable to that of Supplier A*.
2. No residual exonuclease, nicking endonuclease, or RNase detected
Figure 2. Nuclease Contamination Testing of T4 RNA Ligase 2, Truncated KQ
200 units of T4 RNA Ligase 2, Truncated KQ from three independent batches (Yeasen) were incubated with nucleic acid substrates, and reaction products were analyzed by agarose gel electrophoresis. No detectable exonuclease, nicking endonuclease, or RNase activity was observed in any batch, ensuring high reliability and accuracy for sensitive downstream applications.
Documents:
Safety Data Sheet
Manuals
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