[NIPPONGENE] Thermostable Strand-Displacement DNA Polymerase
This product is an enzyme with 5' → 3' DNA polymerase activity and strand displacement activity, synthesizing a new DNA strand by releasing itself from the hydrogen bonds of double-stranded DNA that is used as a template. Thermostable strand-displacement DNA polymerases, which require no dissociation of double-stranded DNA due to their characteristics, can synthesize DNA at a constant temperature without being inhibited by the secondary structure of the DNA.
Features
- 5' → 3' DNA polymerase activity and strand displacement activity
- Able to synthesize DNA at a constant temperature
- Lineup of three enzymes that differ in reaction temperature
- Most suitable for synthesizing DNA strands with high GC content
Use
- Application based on strand displacement activity
(e.g., isothermal amplification)
- Application based on strand displacement activity
Product lineup
| Product name | Optimal temperature | Deactivation temperature*1 | Component | |
|---|---|---|---|---|
| BST DNA Polymerase | 60～65℃ | 80℃, 5 min | 1) BST DNA Polymerase (8units/μl) 2) 10× BST Reaction Buffer (80mmol/l Mg2+) |
1,600units 500μl |
| Csa DNA Polymerase | 60～70℃ | 85℃, 5 min | 1) Csa DNA Polymerase (8units/μl) 2) 10× Csa DNA Polymerase (80mmol/l Mg2+) |
1,600units 500μl |
| 96-7 DNA Polymerase | 50～55℃ | 70℃, 5 min | 1) 96-7 DNA Polymerase (8units/μl) 2) 10× 96-7 DNA Polymerase (95mmol/l Mg2+) |
1,600units 500μl |
*1 Inactivation temperature when enzyme stock solution is directly heat denatured.
≪Bst DNA Polymerase≫ Application for LAMP
LAMP reaction conditions
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Bst DNA Polymerase
Reaction: 65°C, 60 min<Composition of reaction mixture>
Candidatus Liberibacter asiaticus DNA※2 1×106copies FIP 40pmol BIP 40pmol F3 Primer 5pmol B3 Primer 5pmol Loop Primer F 20pmol Loop Primer B 20pmol dNTPs Mixture 1.4mM each 10× Bst Reaction Buffer 2.5 μl Bst DNA Polymerase 8units Total 25μl *2 The LAMP primer set for detection of Candidatus Liberibacter asiaticus was developed by the Kyushu Okinawa Agricultural Research Center of the National Agriculture and Food Research Organization under a project titled "Development of technology to prevent the spread of pesticide-resistant citrus greening disease," one of their projects promoting agricultural forestry and fisheries research using advanced technology.
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＜Lane＞
M：Gene-Ladder Wide 1(Product Number 313-06961)
1 : Company A Bst DNA Polymerase
2 : NIPPONGENE Bst DNA Polymerase＜Note＞
3.0% agarose 21/TAE gel electrophoresis
Ethidium bromide staining
Primer sequence
FIP: 5'-GCATGCCGAGGATCAATGCCTTGCTTAAAGAGCGTGCTACG-3'
BIP: 5'-TATGCCTAATGGCACGGGGGTAAGCTTCATCCGCCTTCGA-3‘
F3 Primer: 5'-TGGGTTAAGTGATGCTGTGG-3'
B3 Primer: 5'-CAACAATATCAGCCCCTGCT-3'
Loop Primer F: 5'-TCTCAACTGTTTCATCAAACCTAGC-3'
Loop Primer B: 5'- CGTGGCGGTTTTTGCTACA-3'
≪Csa DNA Polymerase / 96-7 DNA Polymerase≫ Application for LAMP
LAMP reaction conditions
<Composition of reaction mixture>
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Csa DNA Polymerase
Candidatus Liberibacter asiaticus DNA 1×106copies FIP 40pmol BIP 40pmol F3 Primer 5pmol B3 Primer 5pmol Loop Primer F 20pmol Loop Primer B 20pmol dNTPs Mixture 1.4mM each 10× Csa Reaction Buffer 1× Csa DNA Polymerase 8units total 25μl Reaction: 65°C, 60 min
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96-7 DNA Polymerase
Candidatus Liberibacter asiaticus DNA 1×106copies FIP 40pmol BIP 40pmol F3 Primer 5pmol B3 Primer 5pmol Loop Primer F 20pmol Loop Primer B 20pmol dNTPs Mixture 2.0mM each 10× 96-7 Reaction Buffer 1× 96-7 DNA Polymerase 8units total 25μl Reaction: 65°C, 60 min
Primer Sequence
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FIP: 5'-GCATGCCGAGGATCAATGCCTTGCTTAAAGAGCGTGCTACG-3'
BIP: 5'-TATGCCTAATGGCACGGGGGTAAGCTTCATCCGCCTTCGA-3‘
F3 Primer: 5'-TGGGTTAAGTGATGCTGTGG-3'
B3 Primer: 5'-CAACAATATCAGCCCCTGCT-3'
Loop Primer F: 5'-TCTCAACTGTTTCATCAAACCTAGC-3'
Loop Primer B: 5'- CGTGGCGGTTTTTGCTACA-3' -
<Note>
3.0% agarose 21/TAE gel electrophoresis
Ethidium bromide staining
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<Lane>
M： Gene-Ladder Wide 1(Product Number 313-06961)
1： Strand-displacement DNA polymerase derived from Geobacillus
2： NIPPONGENE Csa DNA Polymerase
3： NIPPONGENE 96-7 DNA Polymerase
≪Csa DNA Polymeras / 96-7 DNA Polymerase≫ Evaluation of thermostability and optimum temperature by RCA method
RCA reaction conditions
<Composition of reaction mixture>
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M13mp18 single strand DNA 20ng Universal Primer 50nM dNTPs Mixture 0.25mM< each Tris-HCl (pH 8.8 at 25℃） 20mM KCl 10mM (NH4)2SO4 10mM MgSO4 2mM Tween 20 0.1% DNA Polymerase 8units total 20μl -
＜Lane＞
M： Gene-Ladder Wide 1(Product Number 313-06961)
D： Template DNA(M13mp18) single strand DNA
Primer Sequence
Universal Primer
5'-GTTTTCCCAGTCACGACGTTGTA-3'
<Note>
3.0% agarose 21/TAE gel electrophoresis
Ethidium bromide staining
RCA(Rolling Circle Amplification) reaction diagram
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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