EB0201 Bst DNA Polymerase

Bst DNA Polymerase

Cat#EB0201

Description

Bst DNA polymerase is Bacillus stearothermophilus DNA polymerase I, a large fragment homolog with improved

isothermal amplification performance and reverse transcription activity.Bst 3.0 DNA polymerase has 5'→3' polymerase activity and strong chain displacement activity towards DNA or RNA templates, but no 5'→3' and 3'→5' exonuclease activity. Compared with Bst DNA polymerase, Bst 3.0 DNA polymerase has strong performance in the amplification of

high concentrations of DNA amplification inhibitors, and the reverse transcriptase activity is also significantly enhanced.

Product Composition

Storage Conditions

Store at -80 ℃. It can be stored at this temperature for one year. Avoid exposure to frequent temperature changes.

Usage

Isothermal amplification (LAMP or RT-LAMP);

DNA Strand Displacement Amplification（SDA）Reaction;

ThermoStable reverse
transciptase (RT) reaction (up to 72 °C);

High amplification capacity in amplification inhibitors or impure samples.

Definition of enzyme activity

1 unit (U) refers to the amount of enzyme required to dope 25 nmol of dNTPs with acid-insoluble material by reacting at 65 °C for 30 min.

Inactivation conditions

Inactivated by reaction at 85 °C for 5 min.

Quality Control

Protein Purity

The purity of Bst DNA polymerase was determined by molecular exclusion high performance liquid phase.

Exonuclease activity

The 20 μL reaction system consisted of 20 U Bst DNA polymerase

and 0.6 μg λ-Hind III were incubated at 37 ℃ for 3 hours. The electrophoretic bands of DNA did not change.

Endonuclease Activity

The 20 μL reaction system consisted of 20 U Bst DNA polymerase

and 0.6 μg λDNA were incubated at 37 ℃ for 3 hours. The electrophoretic bands of DNA did not change.

Ribonuclease residues

The 20 μL reaction system consisted of 20 U Bst DNA polymerase

and 2 μg of RNA were incubated at 37 ℃ for 1 hours. The electrophoretic bands of RNA did not change.

E.coli DNA residues

The E.coli 16 s rDNA gene was amplified in a 20 μL system, using this product as a template, and 30 cycles without amplification.

Experimental Procedures

Configure the LAMP isothermal amplification reaction solution according to the following components, recommended reaction system:

*Primer final concentration recommendation:1.6 μM FIP/BIP，0.4 μM LF/LB，0.2 μMF3/B3.

Mix well, centrifuge for a few seconds, place the reaction in the quantifier at 65 ℃, collect fluorescence signals every minute, set 35-45 cycles. If need to inactivate, the reaction can be set at 85 ℃ for 5-10min.

Bands can be analysed by electrophoresis on a 2% agarose gel if required for the experiment.

Examples of applications

LAMP amplification of N gene plasmid