Product Number: BS07

Shipping and Storage

Transportation under 0°C and storage at -25~ -15°C.

Component

Description

Bst DNA polymerase V2 is derived from Bacillus stearothermophilus DNA Polymerase I, which has 5´→ 3´ DNA polymerase activity and strong chain replacement activity, but no 5´→3´ exonuclease activity. Bst DNA Polymerase V2 is ideally suitable for strand-displacement, isothermal amplification LAMP (Loop mediated isothermal amplification) and rapid sequencing. Well Start Bst DNA polymerase,large fragment is a hot-start version based on Bst DNA polymerase V2 obtained by reversible modification technology, which can inhibit DNA polymerase activity at room temperature, so the reaction system can be operated and formulated at room temperature to prevent non-specific amplification and improve reaction efficiency, and this version can be lyophilized. In addition, its activity is released at high temperatures, so there is no need for a separate activation step.

Application

1. LAMP isothermal amplification;
2. DNA strand single displacement reaction;
3. High GC gene sequencing;
4. DNA sequencing of nanogram level.

Unit Definition

One unit is defined as the amount of enzyme that incorporate 25nmol of dNTP into acid insoluble material in 30 minutes at 65°C.

Heat Inactivation

80℃, 20 min.

Product Number: PC18

Shipping and Storage

Storage at -20℃.

Component

Description

The Klenow Fragment, also known as the Klenow fragment, is a large fragment of Escherichia coli DNA polymerase I (E. coli DNA polymerase I). It retains the 5’→3′ polymerase activity and 3’→5′ exonuclease activity of DNA polymerase I but lacks the complete 5’→3′ exonuclease activity of the full Klenow enzyme. The 3’→5′ exonuclease activity of the Klenow Fragment ensures the fidelity (proofreading) of DNA synthesis.

Application

Fill in at the 5 ‘overhang end of double stranded DNA; Flattening (also known as flattening) the 3 ‘overhang of double stranded DNA; 5 ‘protruding end mark; Random primer method for DNA labeling; Sanger deoxygenation method for DNA sequencing; The synthesis of the second strand of cDNA or site directed mutagenesis reaction.

Features

For adhesive ends with 5 ‘or 3’ protrusions, it can catalyze the generation of flat ends for subsequent flat end connections.

Product Number: PC0302

Shipping and Storage

-20°C

Component

Description

Fast Pfu DNA Polymerase is a fast and highly efficient high fidelity DNA polymerase with 5 ′ -3 ′ DNA polymerase activity and 3 ′ -5 ′ exonuclease activity. This enzyme is modified from other high fidelity enzymes, with strong amplification ability, fast amplification speed, and high fidelity. It overcomes the shortcomings of poor amplification ability, low yield, and slow amplification speed of ordinary Pfu enzymes, greatly shortening the reaction time. This product can be used for long fragment amplification and amplification of various complex templates. The 3 ‘end of the PCR product obtained from amplification does not contain the “A” base and can be directly cloned into a flat end vector. If T/A cloning is required, “A” needs to be added to the end of the PCR product for cloning. This product is suitable for gene cloning, gene site directed mutagenesis, SNP and other amplification experiments.

Definition of Activity

The amount of enzyme required to incorporate 10nmol of deoxyribonucleotides into acidic insoluble substances within 30 minutes at 74℃ is defined as 1 active unit (U).

Quality control

After multiple column purifications, SDS-PAGE analysis showed that its purity was greater than 98%; No exogenous nuclease activity detected; After being stored at room temperature for one month, there was no significant change in activity.

Pfu DNA Polymerase, 5U/μL

Product Number: PC0301

Shipping and Storage

-20°C

Component

Description

This enzyme is a high fidelity heat-resistant DNA polymerase, with a fidelity 8-10 times that of Taq DNA polymerase. This enzyme not only has 5′-3′DNA polymerase activity, but also has 3′-5′exonuclease proofreading activity, which can timely remove nucleotide mismatches in the DNA synthesis chain, greatly increasing the accuracy of base pairing and ensuring the high fidelity of DNA synthesis. This enzyme does not have 5′-3′exonuclease activity.

Source

E. The recombinant strain of Escherichia coli contains genes cloned from Pyrococcus furiosus.

Definition of Activity

The amount of enzyme that can catalyze the polymerization of 10nmol dNTP into a nucleotide fragment by reacting calf thymus DNA as a template at 72℃ for 30 minutes is defined as one active unit (U).

Storage buffer

 [PH8.0] 20mM Tris HCl, 100mM KCl, 0.1mM EDTA, 0.1% NP-40 (v/v), 0.1% Tween-20 (v/v), 1mM DTT, 50% glycerol (v/v).

Reaction buffer

[10 × Pfu Buffer (containing Mg²⁺）] 200mM Tris-HCl（PH 8.8）, 100mM KCl, 160mM(NH4)2SO4，1 % Triton X-100，1 mg/ml BSA, 20mM MgSO4.

Product Number: PH29P

Shipping and Storage

-20℃.

Components

Description

Phi29 Plus DNA polymerase is a protein engineered Phi29 DNA polymerase that is expressed in Escherichia coli and purified and isolated multiple times. Compared with wild-type Phi29 DNA polymerase, it has higher amplification efficiency and sensitivity, and can greatly shorten reaction time. Phi29 plus DNA polymerase has special chain displacement activity and efficient continuous synthesis properties, with strong binding ability to templates. It can continuously synthesize up to 70kb of DNA fragments without dissociating from the template. At the same time, the enzyme has a strong 3´→5´exonuclease correction function, with a fidelity 100 times higher than Taq DNA polymerase.

Application

1. Thermostatic protein prime DNA amplification;
2. Using random primers to amplify DNA;
3. Roll ring replication;
4. Copy extended region.

Features

1. Extremely high infiltration rate;
2. Super strong chain replacement capability;
3. High fidelity;
4. Reaction at moderate temperature;
5. Extremely high sensitivity and amplification efficiency.

Product Number: RNK35H200

Shipping and Storage

-20℃

Components

Description

This product is a recombinant murine RNase inhibitor expressed and purified in Escherichia coli, with a molecular weight of approximately 50 kD. It can specifically inhibit the activity of RNase A, B, and C,Can form a 1:1 complex with RNase, thereby inhibiting its activity. This reaction is reversible, as urea and thiol reagents can dissociate the complex, causing RNase to be renaturated while inhibitors are irreversibly deactivated. When in use, it can be directly added to the reaction solution containing RNA. This product belongs to protein properties and is different from other competitive inhibitors (nucleic acids, inorganic phosphates). It can be easily removed from the reaction system by phenol treatment.

Compared with human RNase inhibitors, murine RNase inhibitors have higher antioxidant activity and are more suitable for experiments with high DTT sensitivity.

This product does not contain ingredients such as glycerol that affect the freeze-drying process, and can be used for the preparation of freeze-drying reaction systems and product design.

Application

This product can be used in any experiment that requires avoidance of RNase interference to prevent RNA degradation, such as:

1. CDNA synthesis reaction.
2. External translation.
3. Polyribosome separation.
4. There is no cellular system transcription in vitro.

Unit definition

The amount of enzyme required to inhibit 50% of 5ng RNase A activity is defined as one activity unit (U). The activity of RNase A was quantitatively determined by inhibiting the hydrolysis of Cyclic2 ‘, 3’ – CMP to generate 3 ‘- CMP

Quality control

After multiple column purifications, SDS-PAGE gel detection showed only a clear and single target band; The qPCR method detects no residual E. coli DNA and no contamination of nucleic acid endonucleases.

Product Number: PC608

Shipping and Storage

-20°C.

Components

Description

Hot Exo- DNA Polymerase purified self weight group E. Coli strain, this enzyme is a natural enzyme obtained through genetic engineering modification.This polymerase is a high-fidelity heat-resistant DNA polymerase with a half-life of 8 hours at 100℃. Suitable for primer extension and high-temperature (72℃) DNA sequencing.

Unit Definition

The amount of enzyme required for the incorporation of 10 nmol of whole nucleotides into acid insoluble precipitates within 30 minutes at 75℃ is defined as 1 active unit (U).

Quality control

The purity of Hot Exo- DNA Polymerase SDS-PAGE is greater than 99%, and there is no activity of endonuclease or exonuclease.

Product Number: LG01

Shipping and Storage

Stored at -20ºC.

Components

Description

T4 DNA ligase can catalyze the phosphodiester bond between the 5 ‘- P and 3’ – OH ends of sticky or flat double stranded DNA or RNA. This catalytic reaction requires ATP as a cofactor. At the same time, T4 DNA ligase can repair single strand defects on double stranded DNA, double stranded RNA, or DNA/RNA hybrids.

Purity: Free of DNA endonucleases, exonucleases, and phosphatases, free of RNAses, meeting the requirements of conventional ligation reactions.

Storage solution:20 mM Tris, pH 7.5, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA and 50% (v/v) glycerol。

10X Ligation Buffer: 400 mM Tris, pH 7.8, 100 mM MgCl₂,100 mM DTT, 10 mM ATP。

Inactivation or inhibition: Incubation at 65ºC for 10 minutes can lead to inactivation of T4 DNA Ligase; When the concentration of NaCl or KCl is greater than 200mM, it strongly inhibits T4 DNA Ligase.

The effect of coating LB plates with T4 DNA Ligase linked products after transformation into competent states is shown in Figure 1.

The LB plate effect obtained by coating the T4 DNA Ligase ligation product onto competent bacteria. A. The carrier after double enzyme digestion was self linked overnight; B. The vector after double enzyme digestion is connected overnight with the fragment to be inserted.

Source

This T4 DNA ligase is expressed by Escherichia coli, and the source of the expressed gene is T4 thermophilic bacteria.

Application

T4 DNA ligase is commonly used for connecting DNA fragments to vectors, linkers, or adapters. It can also be used for defect repair and Ligase mediated RNA detection.

Unit definition

One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of lambda DNA in 30 min at 16ºC in 20µl of the assay mixture containing 50mM Tris, pH 7.5, 10mM MgCl₂, 10mM DTT, 1 mM ATP, 25µg/ml BSA and a 5′-DNA termini concentration of 0.12µM (300µg/ml)。200U is equal to 1 Weiss unit, and based on Weiss unit, this product has a total of 200 units.

Product Number: RI039

Shipping and Storage

Stored at -20ºC, valid for two years.

Components

Description

RNase Inhibitor, Human Placenta,I.e. Ribonuclease Prohibitor, Human Placenta is a recombinant expression of human placental RNase inhibitor in Escherichia coli, which can bind to RNase A, RNase B, RNase C, and human placental RNase in a non competitive 1:1 ratio and inhibit their activity, thereby protecting RNA from degradation by these enzymes.

RNase inhibitor and Human Placenta have extremely strong inhibitory abilities against RNase A, RNase B, RNase C, and human placental ribonuclease, with Ki values as low as about 4×10^-14M. The affinity constant between antibodies and antigens is usually only 10^–6-10^-9M. And the binding between RNase inhibitor and these RNases is very rapid, almost forming complexes with these RNases at the moment of addition to inhibit their enzymatic activity.

RNase inhibitor and Human Placenta cannot inhibit RNase I T1、T2、H、U1、U2、CL3、RNase from Aspergillus、S1 Nuclease、Taq DNA polymerase,M-MLV reverse transcriptase Enzyme activity of SP6, T7, T3 RNA polymerase.

RNase Inhibitor,Human Placenta maintains its RNAse inhibitory activity within the pH range of 5-8, with the highest inhibitory activity observed at pH 7-8. RNase inhibitor requires at least 1mM DTT in the solution to maintain its activity.

This product has His tag on the N-terminus, and if necessary, RNase inhibitor in the solution will be added after the reaction is complete, Human Placenta can be detected by corresponding His antibodies or removed by magnetic beads or nickel columns adsorption.

This product belongs to the same category as Thermo’s RiboLock RNase Inhibitor and Promega’s RNasin Ribonuclease Inhibitor, both of which are recombinant human placental ribonuclease inhibitors with similar inhibitory effects on RNase (refer to Figure 1 and Figure 2).

Figure 1. RNase inhibitor, Comparison of the inhibitory effects of Human Placenta and competitor RNase inhibitor on RNase A enzyme activity. Incubate 5µg of yeast RNA with 0 or 2ng RNase A and 8, 4, 2, 1, or 0U RNase inhibitor in a 100µl reaction system (50mM MOPS, 5mM MgCl₂, pH 6.5) at 37ºC for 15 minutes. After reaction, take 20µl of reaction solution and use 1% agarose gel for electrophoretic analysis. As shown in the figure, this product has a similar inhibitory effect on RNAse activity compared to Competitor’s product. The experimental results obtained under different experimental conditions may vary slightly during actual operation, and the effects shown in the figure are for reference only.

Figure 2. Comparison of the inhibitory effects of RNase inhibitor, Human Placenta, and competitor RNase inhibitor on RNase A enzyme activity. Incubate 2µg of Hela cell total RNA with 0 or 0.5ng RNase A and 20, 10, 5, 2.5, 1.25, 0.625 or 0U RNase inhibitor in a 20µl reaction system (50mM MOPS, 5mM MgCl₂, pH 6.5) at 37ºC for 15 minutes. Immediately after reaction, all samples were electrophoretically analyzed with 1% agarose gel. As shown in the figure, this product has a similar inhibitory effect on RNAse activity compared to Competitor’s product. The experimental results obtained under different experimental conditions may vary slightly during actual operation, and the results shown in the figure are for reference only.

Source

Expressed by Escherichia coli, the source of the expressed gene is the gene encoding the enzyme in human placenta.

Application

Used for protecting RNA from degradation during processes such as cDNA synthesis, in vitro transcription, in vitro translation, and separation and purification of mRNA protein complexes; It can also be used for the identification of specific RNase activity, etc.

Unit definition

The amount of enzyme that can inhibit 50% of the activity of 5ng RNase A is defined as one activity unit.

Activity detection conditions:

100mM Tris-HCl (pH7.5), 1.2mM EDTA, 0.1mg/ml BSA, 100ng/ml RNase A, 0.1mg/ml E.coli [3H]-RNA, 50mg/ml yeast RNA, 8mM DTT。

Purity

Does not contain DNA endonucleases and exonucleases, and does not contain RNAses.

Storage buffer

20mM HEPES-KOH (pH7.5), 50 mM KCl, 5 mM DTT, 50% (v/v) glycerol。

Inactivation or inhibition

Heating at 75ºC for 10 minutes can result in complete deactivation. Heating at 70ºC for 10 minutes will still result in trace amounts of residual activity. Reagents such as SDS and urea that cause protein denaturation, as well as oxidants such as p-chloromercuribenzoate and potassium dichromate, can inhibit the binding of RNase inhibitor to RNAse.

Product Number: PC16F

Shipping and Storage

-20±5°C.

Components

Description

FastStar DNA Polymerase (Glycerol free) is a mixture of glycerol free anti Taq enzyme monoclonal antibodies and Taq DNA Polymerase, suitable for Hot Start PCR.When using Taq enzyme antibodies for PCR amplification, the binding of Taq enzyme antibodies to Taq enzyme inhibits DNA polymerase activity before high-temperature denaturation, which can effectively inhibit non-specific annealing of primers and non-specific amplification caused by primer dimers under low temperature conditions.Taq enzyme antibodies undergo denaturation in the initial DNA denaturation step of PCR reaction, and polymerase activity is restored, achieving a hot start effect. This product does not require special inactivation of Taq enzyme antibodies and can be used under conventional PCR reaction conditions.

FastStar DNA Polymerase (Glycerol free) has 5’→3′ DNA polymerase activity and 5’→3′ exonuclease activity, without 3’→5′ exonuclease activity. The enzyme extends at a rate of 2kb/min and can amplify fragments up to 5kb in length.The amplified PCR product has an “A” base attached to its 3 ‘end, making it suitable for direct T/A cloning. The blocking rate of polymerase activity reaches over 95% at temperatures of 55℃ and below. Heating at 95℃ for 5 seconds can restore DNA polymerase activity.This product has the characteristics of fast extension speed and high amplification efficiency, and is mainly suitable for freeze-drying experiments such as single enzyme freeze-drying, multiple amplification, and DNA sequencing.

Notes

1. After taking out this product from -20℃, please melt it at 4℃ or room temperature. Do not cover it with your hands;
2. After taking out this product, centrifuge it instantaneously before use. This product can be operated at room temperature. If the experimental environment temperature is higher than 25℃ or the experimental time is too long, please place the enzyme on ice;
3. Avoid repeated freeze-thaw of this product, as repeated freeze-thaw (freeze-thaw times≤10 times) may cause a decrease in product performance. This product can be stored for a long time at -20±5℃;
4. This product is recommended for small portion packaging.

Quality Control

1. Protein purity: The purity detected by HPLC is close to 99%;
2. Detection of Exonuclease Residues:10U of proenzyme and 0.6μg λ-Hind III was incubated at 37℃ for 16 hours, and the electrophoresis bands of DNA did not change.
3. Endonuclease residue detection:10U of proenzyme and 0.6μg Supercooled pBR322 DNA was incubated at 37℃ for 4 hours, and the electrophoresis bands of the DNA did not change.
4. RNase residue detection:10U of proenzyme and 1μg HeLa cell total RNA at 37℃ for 1 hour, and the electrophoresis band of RNA remains unchanged.