Bst DNA Polymerase is an enzyme derived from the large fragment of Bacillus stearothermophilus DNA Polymerase I. It contains 5´- 3´ DNA polymerase activity and strong strand displacement activity but lacks 5´- 3´ exonuclease activity. The strong strand displacement activity enables Bst DNA Polymerase to synthesize DNA at a constant temperature making it an ideal enzyme for Loop-Mediated Isothermal DNA Amplification (LAMP). Two optimized buffers are available that are formulated to deliver the best amplification speed, yield, salt tolerance and sensitivity. The Inhibitor-Tolerant Bst Buffer is specifically designed for high performance when using crude samples such as sputum or saliva.
Bst DNA Polymerase & Optimized Reaction Buffers for Isothermal Assays
APPLICATION KEY
Environmental Human Diagnostics Vet Health
Food Testing Water Testing DNA Barcoding
Product
Cat No.
Volume
Bst DNA Polymerase is an enzyme derived from the large fragment of Bacillus stearothermophilus DNA Polymerase I. It contains 5´- 3´ DNA polymerase activity and strong strand displacement activity but lacks 5´- 3´ exonuclease activity. The strong strand displacement activity enables Bst DNA Polymerase to synthesize DNA at a constant temperature making it an ideal enzyme for isothermal amplification, such as HDA, MCA and Loop Mediated Isothermal Amplification (LAMP). Two optimized buffers are available that are formulated to deliver the best amplification speed, yield, salt tolerance and sensitivity. The Inhibitor-Tolerant Bst Buffer is specifically designed for high performance when using crude samples such as sputum or saliva.
Enzyme Buffers
2 mL 100 mL 5 mL 100 mL 1 mL 20 mL 20 mL* 80 μL 800 μL 800 μL* 5 mL 100 mL 5 mL 100 mL
Inhibitor-Tolerant Bst Buffer
MDX019
Bst Reaction Buffer, 10x
MDX076
Polymerases
Bst DNA Polymerase
MDX012
High Conc. Glycerol-Free Bst MDX018
Dilution Buffers
Product Highlights
Enzyme Dilution Buffer, 1x MDX078
• Optimized for Isothermal Amplification • Reaction buffer and dilution buffers sold separately for maximum flexibility • Inhibitor-Tolerant Bst Buffer is ideal for using crude samples such as sputum or saliva
Enzyme Dilution Buffer (10X) Glycerol free
MDX080
*Enzyme only
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High Performance Buffer
Inhibitor-Tolerant Bst Buffer ( MDX019 )
Data Highlights
Efficient amplification over different GC content and in the presence of sputum LAMP amplification of B. pertussis (57% GC), S. aureus (33% GC), Epstein-Barr Virus (59% GC) and M. tuberculosis (65% GC) in the presence of 20% artificial sputum medium. Inhibitor- Tolerant Bst Buffer (MDX019, dark orange) shows faster time-to-result (TTR) compared to a Standard Bst Buffer (blue) .
High reaction efficiency in the presence of increasing concentrations of saliva LAMP amplification of hBRCA1 gene from human genomic DNA spiked into human saliva with Inhibitor-Tolerant Bst Buffer (MDX019, dark orange) and Standard Bst Buffer (blue) . Inhibitor-Tolerant Bst Buffer shows no loss of activity with increasing concentrations of saliva.
Saliva (0-30%) final reaction concentration
Respiratory pathogen detection in presence of sputum
60
1 2 3 4 5 6 7 8 9 10
50
40
30
20
10
0
0
B. pertussis
S. aureus
Epstein Barr Virus
M. Tuberculosis
0
2%
10% 20% 30%
Inhibitor-Tolerant Bst Buffer
Standard Bst Buffer
Inhibitor-Tolerant Bst Buffer
Standard Bst Buffer
Improved tolerance to Universal Transport Medium (UTM)
T. vaginalis DNA was spiked into UTM and added at final concentrations of 5%, 20% or 40% to reactions containing Inhibitor-Tolerant BST Buffer (MDX019, dark orange) or a Standard Bst Buffer (blue) . Reactions with Inhibitor-Tolerant Bst Buffer demonstrated better activity at high concentrations of UTM compared to reactions using only a Standard Buffer.
5% UTM
20% UTM
40% UTM
Higher reaction specificity LAMP amplification of 100 input copies of DNA template from B. pertussis, S. aureus , Epstein-Barr Virus and M. tuberculosis , along with non-template controls (NTC) using Inhibitor-Tolerant Bst Buffer (MDX019, dark orange) or a Standard Bst Buffer (blue) . Reactions containing Inhibitor-Tolerant Bst Buffer showed no NTC amplification (brown) or delayed amplification compared to reactions containing a Standard Bst Buffer.
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Bst Reaction Buffer, 10x ( MDX076 )
Data Highlights
Exhibits High Tolerance to Salt Inhibition LAMP reactions were performed using Bst Reaction Buffer/Bst Polymerase (MDX076/MDX012, orange) or a competitor Bst Polymerase and buffer (gray) using increasing amounts of salt (potassium chloride). The time to results (TTR) illustrate that Bst Reaction Buffer performs well in the presence of high salt concentrations which can inhibit LAMP reactions.
Bst DNA Polymerase
Standard Bst Buffer
Enables Fast Polymerization LAMP reactions were performed using Meridian’s Bst Reaction Buffer/Bst DNA Polymerase (MDX076/MDX012, orange) and compared against three competing Bst enzymes/buffers [Supplier A (blue) , Supplier B (green) and Supplier C (purple) ]. Meridian’s Bst buffer/enzyme consistently reached the fastest reaction threshold times across three different targets (BRCA1 tumor suppressor gene, CFTR and S. stercoralis ).
A/ BRCA1 tumour suppressor gene
B/ Cystic Fibrosis Transmembrane Receptor (CFTR)
C/ Strongyloides stercoralis (Human pathogenic parasite)
Time [60 s interval]
Time [60 s interval]
Time [60 s interval]
High Stability Under High Temperature or Repeated Freeze/Thaw Bst Reaction Buffer/Bst DNA Polymerase (MDX076/MDX012) was left at (A) +4 ° C, ambient (room temperature) or +37 ° C for 4 hours, 24 hours or 1 week and (B) subjected to 5, 20 and 30 rounds of fast freeze/thawing, between -80 ° C and 0 ° C. The results illustrate that the Bst Mix MDX076/MDX012 is very stable, and able to withstand temperatures up to +37 ° C for a 1 week, or up to 30 freeze/thaw cycles without losing activity. Similar results were obtained with Bst Standard Buffer/ High-Conc. Glycerol-Free Bst Polymerase (MDX076/MDX018) under the same testing conditions. Data is available on request.
B/
A/
12.0 11.0 10.0
12 11 10
9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0
9 8 7 6 5 4 3 2 1 0
+4 o C
Ambient Storage Temperature
+37 o C
5
20 Freeze-thaw Cycles
30
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