Cat.No.：D21011A
Price: ￥
Specifications：
T5 nucleic acid exonuclease is a 5'→3' directionally multifunctional nucleic acid exonuclease derived from bacteriophages, and it also possesses single-stranded DNA endonuclease activity. This enzyme can efficiently digest linear and nicked single-stranded DNA and double-stranded DNA, but does not degrade supercoiled intact double-stranded DNA.
Thanks to its unique substrate recognition characteristics, T5 nucleic acid exonuclease can precisely digest linear residual fragments and nicked DNA, while completely preserving supercoiled plasmids. It is a core tool enzyme for Gibson seamless assembly, plasmid purification, and DNA fragment modification experiments, and is suitable for various molecular cloning experimental systems.
1. Specificity of substrate is strong
It specifically degrades linear, looped, and single-stranded DNA, without damaging intact supercoiled plasmids, and can efficiently enrich high-purity supercoiled DNA templates.
2. Extremely high enzyme purity
After being processed by recombinant protein purification technology, there is no contamination from other enzymes, and no residual endonucleases, exonucleases, or RNase. The experimental background is clean and the results are highly reproducible.
3. Stable degradation efficiency
By optimizing the recombinant expression process, the enzyme activity is high, the reaction rate is stable, and DNA end digestion can be completed quickly, suitable for batch assembly and high-throughput cloning experiments.
4. Complete supporting system
Combined with the dedicated 10× reaction buffer, the reaction system has good compatibility and high adaptability. No complex debugging is required, and it can be used directly, significantly reducing experimental errors.
| Cat. No. | Component | Specification | Quantity |
| B28011A | T5 Exonuclease | 1000 U | 1 |
| B28012A | 10×T5 Reaction Buffer | 3 mL | 1 |
Storage conditions: Store at -20℃ in a sealed container for a long term. Avoid repeated freezing and thawing. The enzyme activity can be stably retained for a long time.
Transport conditions: Transport throughout the process under low-temperature cold chain (≤0℃) to ensure that the enzyme activity is not lost during transportation.
1. Gibson seamless cloning assembly: Utilizes mainstream constant-temperature assembly core enzymes to digest DNA fragments at their 5' ends to generate complementary sticky ends, enabling efficient and seamless assembly of multiple fragments.
2. High-purity plasmid preparation: Specifically degrades linear DNA and removes fragmented plasmid impurities, effectively enriching and purifying supercoiled plasmids, thereby improving sequencing and transfection quality.
3. DNA fragment modification and purification: Removes residual linear amplification fragments, incomplete ligation products, and redundant single-stranded DNA, optimizing the background of molecular cloning experiments.
4. Various molecular biology experiments: Adapts vector modification, fragment trimming, and optimization of cloning systems for both routine and advanced molecular experiments.
What types of DNA can the T5 exonuclease act upon?
Linear double-stranded DNA (dsDNA) or single-stranded DNA (ssDNA) are acceptable. If the target DNA has a gap, digestion will start from that position. However, this product cannot act on supercoiled double-stranded DNA.
What are the main differences between T5 exonuclease and λ exonuclease?
The λ exonuclease strictly begins digestion from the 5' end of the double-stranded DNA and prefers the phosphorylated 5' end. It has no activity on single-stranded DNA. The T5 exonuclease not only can start digestion from the 5' end of the linear double-stranded DNA, but also can start digestion from the cleavage or nick of circular DNA, and has endonuclease activity for single-stranded DNA, with a wider substrate range.
How to terminate the reaction when using T5 exonuclease to degrade DNA?
EDTA can be added to the reaction system to a final concentration of ≥ 11 mM to chelate the Mg²⁺ ions in the reaction system, or DNA loading buffer containing SDS can be added to achieve a final concentration of SDS of 0.08%.
