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EB0106 Bsu DNA Polymerase

Cat. No. EB0106
Source /
Appearance Liquid
Endotoxin /
Purity /
Consult Immediately
Detail

Bsu DNA Polymerase

Cat#EB0106

Description

Bsu DNA Polymerase is derived from Bacillus subtilis and has a molecular weight of about 68 kDa. Large fragments of Bsu DNA polymerase retain 5 '→3' polymerase activity, but lack 5 '→3' exonuclease activity, and naturally lack 3 '→5' exonuclease activity. The optimum reaction temperature of the enzyme is 37 ℃, and it has strong chain replacement activity, which is suitable for DNA isothermal amplification and cDNA double chain synthesis.

Product Composition

Name

Size

Bsu DNA Polymerase

10 mg/mL, 500 μL

Storage Conditions

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

Store at -80 ℃. Avoid exposure to frequent temperature changes.

Usage

The enzyme can be used for fluorescent RT-RPA or RPA, depending on the optimization degree of primer screening and detection means, the detection limit is 10-100 copies/test, and the reaction time is not more than 15 minutes.

Stability

When placed at -80 ℃ for 3 months, the enzyme application activity did not decrease significantly (data update).

Freezing and thawing for 5 times showed no significant decrease in enzyme activity.

The enzyme application activity did not decrease significantly after being placed at 4 ℃ for 21 days (data update).

After lyophilization, the enzyme application activity did not decrease significantly.

75 ℃, 20 minutes of inactivation.

Quality Control

Protein Purity

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

Endonuclease Activity

The 50 μL reaction system consisted of 10 μL single-stranded DNA-binding protein and 500 ng circular plasmid DNA. After incubation at 37 ℃ for 4 h, the endonuclide activity was detected by agar-gel electrophoresis.

Exonuclease activity

The 50 μL reaction system consisted of 10 μL DNA polymerase and 500 ng double-stranded DNAcircular plasmid DNA. After incubation at 37 ℃ for 4 h, the activity of endonuclide was detected by agarose gel electrophoresis.

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.

Application Example

Enzyme Activity Assays

Bsu1.pngBsu2.png

Figure 1. Amplification sensitivity of different types of templates. The RT-RPA reaction system consisted of recombinaserecombinant enzyme, recombinaserecombinant enzyme helper protein, single-stranded DNA binding protein, DNA polymerase, reverse transcriptase 4.0, exonuclease III, RT-PRA buffer, PEG and trehalose. The reaction was performed in a qPCR apparatus at 42 ℃ for 25 minutes. (A) An RNA virus template; (B) A DNA plasmid template.

Enzyme stability test

Bsu4.pngBsu3.png


Figure 2.   .Stability of DNA polymerase. The DNA polymerase was freeze-dried in storage solution (A) and placed at 4 ℃ for 21 days (B). The stability of the enzyme was determined by RT-RPA reaction system. Template: RNA virus, template amount: 104 and 103 copies/reactions.