Product Number: EN03

Shipping and Storage

-20℃.

Components

Description

Exonuclease III can gradually cleave single nucleotides along the 3 ‘→5’ direction of double stranded DNA. Each reaction only removes the last few nucleotides to produce progressive deletion of double stranded DNA. This enzyme can cleave double stranded DNA, producing single stranded gaps, but is inactive against single stranded DNA. DNA with flat or 3 ‘recessed ends is usually the most suitable substrate. On the contrary, the 3 ‘protruding end is resistant to cleavage by the enzyme, and the degree of antagonism varies with the length of the 3’ protruding end. Protrusions of four bases or longer cannot be cleaved at all. This characteristic can result in unidirectional deletions in linear DNA molecules with one end being a resistant site (3 ‘protruding end) and the other end being a sensitive site (flat or 5’ protruding end). In addition, compared with BAL 3 Nuclease, this enzyme has lower base specificity. For example, at positions rich in GC, due to the lower probability of reaction stoppage, it can be used for DNA Deletion production.

This product is obtained by expressing the Exonuclease III gene (E. coli) in Escherichia coli and purifying and isolating it multiple times.

Application

1. Site directed mutation;
2. Preparation of chain specific probes;
3. Preparation of single stranded substrates for deoxygenation sequencing.

Product Number: DB005

Shipping and Storage

-20℃.

Components

Description

This product is a Cre recombinase obtained by expressing the plasmid encoding the bacteriophage P1 Cre protein in Escherichia coli and purifying and isolating it multiple times. This enzyme does not require energy cofactors, and Cre – mediated recombination quickly reaches equilibrium between the substrate and the reaction product.

Cre recombinase is a type I topoisomerase of bacteriophage P1, catalyzing site-specific recombination of DNA between loxP sites. The loxP recognition element is a 34bp sequence with two 13bp inverted repeat sequences at both ends and an 8bp spacer in the middle for directional purposes. The recombinant product varies depending on the position and relative direction of the loxP site, and two DNA containing a single loxP site will be fused. The DNA between two forward repeating loxP sites will be cleaved in a circular form, while the DNA sequence between two reverse loxP sites will be flipped.

Application

1. DNA cleavage between two loxP sites;
2. Fusion of DNA molecules containing loxP sites;
3. Flipping of DNA sequences between loxP sites.

Product Number: DB004

Shipping and Storage

Low temperature transportation; Shelf life of one year at -20℃.

Components

Description

The DNA topoisomerase I produced by our company is a type I eukaryotic topoisomerase derived from the smallpox virus. The DNA Topoisomerase I has DNA Relaxing activity, which can cause double stranded covalently closed circular positive or negative supercoiled DNA to undergo supercoiling, thereby forming double stranded circular DNA molecules with fewer positive or negative supercoils. Vaccinia DNA Topoisomerase I can also cause double stranded DNA to form knots or undo knots, resulting in complementary single stranded circular DNA becoming double stranded circular DNA. In addition, Vaccinia DNA Topoisomerase I can specifically recognize the 5 ‘-… (C/T) CCTT… -3’ sequence in double stranded DNA, and can open a phosphodiester bond on one strand after the 5 ‘-… (C/T) CCTT↓sequence. The energy released by breaking the phosphodiester bond can catalyze the 3’ phosphate group at the DNA incision to form a covalent ester bond with the 274th tyrosine (Try) hydroxyl group of topoisomerase, thus forming a covalent complex between DNA and the enzyme. The covalent ester bond formed simultaneously can be attacked by the 5 ‘hydroxyl group of the cleaved single stranded DNA, and without the need for ATP and DNA ligase, the previously formed phosphodiester bond can be restored, that is, the DNA can be reconnected while releasing Vaccinia DNA Topoisomerase I.

Application

At present, DNA Topoisomerase I (vaccine virus) is often used as a novel tool enzyme for DNA recombination and ligation, for the ligation, defect repair, and linker ligation of DNA vectors and recombinant fragments.

Features

1. Features: DNA supercoiling and specific opening of phosphodiester bonds.
2. Source: Escherichia coli strains carrying the Vaccinia virus topoisomerase gene; Its molecular weight is about 37KDa;
3. Enzyme activity definition: The amount of enzyme required to completely unwind 1µg of pUC19 carrier after incubation at 37℃ for 30 minutes is defined as one enzyme activity unit.
4. Purity and concentration: SDS-PAGE detection shows a purity of ≥95%; Endogenous nucleic acid residue<1pg/μL (qPCR detection); 10U/μL。
5. Inactivation or inhibition: Heating at 80 ° C for 20 minutes can fully inactivate DNA Topoisomerase I.
6. Enzyme storage buffer：50mM Tris-HCl，1mM DTT，0.1mM EDTA，0.1 M NaCl，0.1%Triton X-100，50%glycerol，pH 7.5.
7. 10х Reaction buffer：500mM Tris-acetate, 1M NaCl, 25 mM MgCl_2, 1mM EDTA，pH 7.5.

Product Number: DB003

Shipping and Storage

-20℃.

Components

Description

This enzyme is obtained by expressing the phosphatase gene isolated from the Antarctic cold loving strain TAB5 in Escherichia coli and purifying it multiple times. Phosphatases can catalyze the removal of 5 ‘phosphate groups at the ends of DNA or RNA. After treating the vector DNA with thermosensitive phosphatase during cloning, its fragment lacks the necessary 5 ‘phosphate terminus for ligase, so the vector cannot perform self ligation, which can greatly reduce the background of vector self ligation. This enzyme can act on the 5 ‘protruding end, concave end, and flat end.

Application

1. Remove the phosphate group at the 5 ‘end of the carrier to reduce the background of self connection;
2. Remove residual dNTP and pyrophosphate from PCR products;
3. Remove the phosphate group at the 5 ‘end of the DNA probe and prepare the optimal substrate for phosphorylation labeling at the 5’ end;
4. Protein dephosphorylation.

Product Number: TE035

Shipping and Storage

-20℃.

Components

Description

T5 Exonuclease is encoded by the D15 gene of T5 bacteriophage and can degrade DNA along the 5 ‘-3’ direction. It can be digested from both the 5 ‘end and the cleavage or gap of linear or circular double stranded DNA, but cannot degrade supercoiled DNA. The single stranded endonuclease activity of this enzyme can be inhibited by low concentrations of Mg^2+less than 1mM.

Application

1. Degradation of ssDNA and dsDNA;
2. Degradation of plasmid DNA with cleavage gaps;
3. When degrading single stranded and double stranded DNA, it can preserve the supercoiled plasmid DNA.

Features

1. Efficient and sensitive: Short degradation time, high degradation rate for single and double stranded DNA fragments, and sensitive response.
2. High quality: fully meets the experimental requirements for degrading single and double stranded DNA fragments.
3. Unique formula of 10 × Reaction Buffer: makes substrate degradation reaction more stable, sensitive, and efficient.

Product Number: TE101

Shipping and Storage

Storage at -20±5℃.

Component

Description

This product is a protein cloned from the TelN Protelomerase gene expression of phage N15. It can specifically recognize the telRL sequence (56bp) on dsDNA, cleave dsDNA, and form a covalently closed end at the cleavage site, effectively transforming circular DNA into linear DNA with a closed end. The closed ended linear DNA generated by TelN Protelomerase treatment has stable performance, long half-life, and only introduces two 28 bp short sequences except for necessary sequences. It can encode long, complex, or unstable DNA sequences, does not contain bacterial sequences, and has a strong expression profile.

Application

This product can be used in fields such as DNA vaccine development, mRNA vaccine development, virus vector preparation, DNA data storage, etc.

Features

This product has strong specificity and can specifically cleave the recognized DNA sequence (telRL) to form a covalently closed end. The recognition sequence is as follows:

Unit definition

1 unit refers to the amount of enzyme required to cleave 0.5μg BsaI linearized plasmid (313fmol telRL recognition site) in a 50μL 1 × TelN Reaction Buffer reaction buffer system at 30℃ for 30 minutes.