Bst DNA Polymerase

Bst DNA Polymerase & Optimized Reaction Buffers for LAMP 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 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.

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

Product Highlights

Dilution Buffers

• Optimized for Loop-Mediated Isothermal DNA Amplification (LAMP) • 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, 1x MDX078

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

Ordering information: USA 5171 Wilfong Road Memphis, Tennessee 38134 Fax: +1 901-333-8223 Toll Free: +1 800 327 6299

Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.MeridianLifeScience.com

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