T5 Exonuclease

Cat. No.: T750876

GRADE & PURITY Suitable for molecular biology, BioReagent, EnzymoPure™, Exonuclease, ≥95%(SDS-PAGE), ≥10 U/μL

Overview

Storage buffer solution: 50mM Tris-HCl, 100mM NaCl, 1mM DTT, 0.1mM EDTA, 50% Glycerol, 0.1% Triton X-100, pH 7.5, 25ºC
Product Introduction:
The T5 Exonuclease produced by Aladdin, also known as T5 nuclease, is a nuclease that degrades double stranded or single stranded DNA in the 5 '→ 3' direction. T5 Exonuclease can be digested starting from the 5 'end of single stranded or double stranded DNA, as well as from the gap or nick of linear or circular double stranded DNA. T5 Exonuclease is unable to degrade supercoiled double stranded DNA, and its activity in degrading single stranded DNA can be inhibited by reducing the Mg "2+in the reaction buffer to below 1mM. Based on these characteristics, T5 Exonuclease is commonly used for Gibson Assembly. Gibson assembly is a technique that effectively connects multiple overlapping sequence fragments under constant temperature conditions. Its basic principle can be summarized as three steps: (1) T5 nuclease digests from the 5 'end of the DNA fragment, producing complementary single stranded 3' ends (overhangs) and promoting annealing of the complementary ends; (2) DNA polymerase fills gaps in annealed fragments; (3) DNA ligase connects the assembled nick sites to obtain a complete double stranded DNA.
Purpose:
Commonly used for Gibson assembly; Degradation of linear single stranded, double stranded DNA or truncated plasmid DNA; Removing incomplete ligation products from connected circular double stranded DNA; Degradation of linear and truncated plasmid DNA to obtain high-purity supercoiled plasmid DNA; Remove the denatured plasmid DNA generated during the process of extracting plasmids using alkaline splitting method; Improve the transfection efficiency of small-scale extraction plasmid cDNA libraries.
Definition of Activity
1. The active unit (U) is the amount of enzyme required to generate 1 nmol acid soluble deoxyribonucleotide from double stranded DNA, with a total reaction volume of 50 µ l and a reaction time of 30 minutes at 37 º C.
matters needing attention:
T5 Exonuclease is a selective nuclease for DNA substrates, exhibiting varying reactivity towards different DNA substrates. Therefore, when digesting specific substrates, it is important to control the enzyme dosage and reaction time appropriately. This product is only for scientific research by professionals and should not be used for clinical diagnosis or treatment, food or medicine, or stored in ordinary residential areas. For your safety and health, please wear lab coats and disposable gloves when operating.

Specifications

Product Name

T5 Exonuclease

Synonyms

T5 Exonuclease

Grade

BioReagent, Exonuclease, Suitable for molecular biology, EnzymoPure™

Specifications & Purity

BioReagent, Suitable for molecular biology, EnzymoPure™, Exonuclease, ≥95%(SDS-PAGE), ≥10 U/μL

Molecule Type

Enzyme

Storage and Shipping

Concentration

≥10 U/μL

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20℃ long term (24 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Unit definition

One unit is defined as the amount of enzyme required to produce 1 nmol of acid soluble deoxyribonucleotide from double-stranded DNA in 30 minutes at 37 ℃ in a total reaction volume of 50μl.