T7 Exonuclease, encoded by gene 6 of bacteriophage T7, is a highly processive 5'→3' exonuclease exhibiting robust enzymatic activity. This enzyme specifically catalyzes the double-stranded DNA (dsDNA) or RNA-DNA heteroduplexes, initiating hydrolysis from either 5'-phosphorylated or 5'-hydroxyl termini. The enzymatic reaction proceeds via the sequential release of oligonucleotides and mononucleotide products.
Notably, T7 Exonuclease demonstrates unique substrate versatility, capable of initiating digestion not only at terminal 5' ends but also at internal nicks or gaps within duplex structures. Importantly, the enzyme exhibits strict substrate specificity, showing no detectable activity against either single-stranded or double-stranded RNA molecules. This characteristic makes it particularly valuable for molecular biology applications requiring selective nucleic acid processing.
Figure 1. Schematic illustration of the enzymatic activity of T7 Exonuclease
Owing to these distinctive properties, T7 Exonuclease plays a pivotal role in several critical applications in molecular biology research, including seamless cloning, single-stranded DNA template preparation, random mutagenesis library construction, and digestion of uncyclized linear templates.
To meet researchers' demand for high-quality enzymes, Yeasen has newly launched T7 Exonuclease. This product exhibits robust digestive activity and undergoes stringent quality control, being free from contaminating nickase and RNase activities, thereby ensuring unimpaired experimental results.
Characterization of Yeasen T7 Exonuclease (Cat#14272)
- Excellent enzymatic digestion performance
Figure 2. Performance evaluation of DNA digestion and heat inactivation characteristics in T7 Exonuclease preparations of Yeasen and Supplier A*
Note: Under the reaction conditions of 25°C for 30 minutes, the digestion efficiency of 10 U of Supplier A* T7 Exonuclease and three batches of Yeasen T7 Exonuclease on 1 μg of calf thymus DNA was compared. Additionally, the thermal inactivation efficacy of T7 Exonuclease was evaluated after incubation at 75°C for 20 minutes and 80°C for 10 minutes. The results demonstrated that all three batches of Yeasen T7 Exonuclease effectively digested the substrate, and both 75°C for 20 minutes and 80°C for 10 minutes successfully inactivated the T7 Exonuclease.
- No Detectable Nickase or RNase Residues
Figure 3. Analysis of Nickase and RNase Residual Activities in Yeasen T7 Exonuclease Preparations
[Note]: T7 Exonuclease was incubated with nucleic acid substrates respectively, and the banding patterns were analyzed by agarose gel electrophoresis. The experimental results demonstrated that no residual nickase (100 U) or RNase (10 U) activities were detected in any of the three batches of Yeasen T7 Exonuclease.
Ordering Information
|
Product Name |
Packaging Specifications |
Catalog No. |
|
T7 Exonuclease (10 U/μL) |
1000 U / 5000 U |
14272ES |
Extended Reading
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References
[1] Holland E G, Acca F E, Belanger K M, et al. In vivo elimination of parental clones in general and site-directed mutagenesis[J]. Journal of immunological methods, 2015, 417: 67-75.
[2] Wang M, Fu Z, Li B, et al. One-step, ultrasensitive, and electrochemical assay of microRNAs based on T7 exonuclease assisted cyclic enzymatic amplification[J]. Analytical Chemistry, 2014, 86(12): 5606-5610.
[3] Tabor S, Richardson C C. Selective inactivation of the exonuclease activity of bacteriophage T7 DNA polymerase by in vitro mutagenesis[J]. Journal of Biological Chemistry, 1989, 264(11): 6447-6458.