Features/Applications:
FAST 2.0 Bst Mastermix is an extraordinary candidate for isothermal amplifications:
- DNA strand displacement amplification (SDA)
- Cross priming amplification (CPA)
- Rolling-circle amplification (RCA)
- Loop-mediated isothermal amplification of DNA
(LAMP)
- Reverse transcription isothermal multiple-self-
matching-initiated amplification (RT-ISMA)
- Polymerase chain displacement reaction (PCDR)
- Sequencing of very low amounts of DNA template
General benefits of isothermal amplification:
- More resistant in Plant tissue of blood samples to
inhibitors
- Very fast screening of many DNA samples
- Inexpensive and simple method
- Extreme sensitive detection
Description:
FAST 2.0 Bst Mastermix (2x), genetically improved Bacillus stearothermophilus cloned to E. Coli in a ready-to-use Mastermix, for next generation of isothermal DNA amplification.
The Mastermix allows the detection within up to 5 to 10 minutes (when using an extra primer pair) and thus it is 2 to 3 times quicker than other Bst DNA Polymerases.
The optimized FAST 2.0 Bst Mastermix amplification results can be compared with about 28-31 cycles in a standard PCR-Cycler Platform. The FAST 2.0 Bst Mastermix offers high strand displacement capability. The optimal temperature range is between 60°C and 65 °C.
Method of detection:
We recommend to use the fluorescent DNA stain EvaGreen to add in the reaction Mix or to use the ready to use FAST Mastermix 2.0 Bst with Evagreen Cat.-No. S650 or S660 (Mastermix 2.0 Bst with EvaGreen and ROX)
Content:
Fast 2.0 Bst Polymerase, dNTPs, reaction buffer, glycerol, EvaGreen as intercalating dye, stabilizer and enhancer.
READY-to-use. The Mastermix is 2x concentrated
The Mastermix is available with high ROX content as normalization dye.
Concentration: 2 x
Storage condition: @ -20°C; for short term (up to 10-12 weeks) at +4°C
Shipping: on blue ice
Protocol for 50 µl reaction volume
Component |
Concentration final |
Volume |
FAST 2.0 Bst Mastermix Evagreen dye with or without ROX; (10x) |
1x | 25 µl |
Primer Mix (10x) | 1x | 5 µl |
Template DNA | max. 500 ng | X µl |
PCR-Water | up to 50 µl |
BST Mastermix is available with Evagreen product-code S650, or Evagreen and High ROX product-code S660
Applications:
FAST 2.0 Bst Mastermix is an extraordinary candidate for isothermal amplifications:
- DNA strand displacement amplification (SDA)
- Cross priming amplification (CPA)
- Rolling-circle amplification (RCA)
- Loop-mediated isothermal amplification of DNA (LAMP)
- Reverse transcription isothermal multiple-self-matching-initiated
amplification (RT-ISMA)
- Polymerase chain displacement reaction (PCDR)
- Sequencing of very low amounts of DNA template
Final price excl. shipping costs3
Applications:
FAST 2.0 Bst Mastermix is an extraordinary candidate for isothermal amplifications:
- DNA strand displacement amplification (SDA)
- Cross priming amplification (CPA)
- Rolling-circle amplification (RCA)
- Loop-mediated isothermal amplification of DNA (LAMP)
- Reverse transcription isothermal multiple-self-matching-initiated
amplification (RT-ISMA)
- Polymerase chain displacement reaction (PCDR)
- Sequencing of very low amounts of DNA template
Final price excl. shipping costs3
1. A LAMP Primer Mix can be prepared with all 4 or 6 (with Loop) primers. A 10X Primer Mix should contain: 16 µM FIP, 16 µM BIP, 2 µM F3, 2 µM B3, 8 µM LoopF, 8 µM LoopB in TE or water
2. Reactions should be setup on ice.
3. If analyzing via agarose gel electrophoresis or other method requiring opening LAMP reaction vessels, setup secondary analysis area and equipment to avoid contamination.
4. Running a no-template control is strongly recommended to ensure amplification specificity.
5. If optimization is desired, try titrating Mg2+ (4–10 mM final) or Bst DNA Polymerase, Large Fragment (0.01–0.2 U/µl), or changing reaction temperature (50–68°C).
6. dNTP should be used with dTTP, not dUTP
7. We recommend using LAMP primer design software such as Primer Explorer (http://primerexplorer.jp/e/).
Bst DNA Polymerase