Saphir Bst Turbo GreenMaster Lyophilisate

For general laboratory use.

Envio: shipped at ambient temperature

Condições de armazenamento: store at ambient temperature
Store in an aluminium-coated bag or on a dry place.
Lyophilisates may hydrate at humidity levels >70 % when sealing is opened.

Validade: 6 months in sealed packaging

Descrição:
Saphir Bst Turbo GreenMaster Lyophilisate is designed for isothermal amplification of DNA. The mix is based on a genetically enhanced Bst polymerase of the next generation. The mixe is the ideal choice for ultra-fast and robust amplification of DNA at constant temperature (60 to 65 °C). The enzyme shows high strand displacement activity and generates an amplification factor of up to 109 which is comparable to approx. 30 cycles in a PCR assay. The polymerase is 2-3x faster compared to Saphir Bst Polymerase (#PCR-389/#PCR-388/#PCR-387) and allows detection of a target gene within 5-10 minutes.

Contente:
Saphir Bst Turbo GreenMaster Lyophilisate
Saphir Bst Turbo Polymerase, dNTPs, reaction buffer, Green DNA intercalator dye, additives and stabilizers

PCR grade water

Handling
Saphir Bst Turbo GreenMaster Lyophilisate is delivered is delivered in 8-tube strips or 96-well plates preloaded with a complete master mix in a dry, room temperature stable format. The lyophilisate combines highest performance with convenience of use and stability. There is no need for freezing, thawing or pipetting on ice. The few remaining pipetting steps minimize the risk of errors or contaminations.
Each vial contains all components (except primers and template) required for a 20 μl LAMP assay.
To perform the Assay, only fill up the vials with a primer mix and add DNA template.
The lyophilisate can also be used with ROX reference dye in PCR instruments that are compatible with the evaluation of the ROX signal. In this case, the ROX dye (#PCR-351) should be added as 1x concentration to the PCR reaction.

Assay design
Isothermal amplification is an extremely sensitive detection method and care should be taken to avoid contamination of set-up areas and equipment with DNA of previous reactions. A problem may be amplification in no-template controls due to carry-over contamination or amplification of unspecifically annealed primers or primer dimer formations.

Primer design
Typically, 4 different primers are used to identify 6 distinct DNA regions allowing the specific amplification of a target gene. An additional pair of primers further accelerates the amplification allowing to cut down the total detection time to 10-20 min.
The manual design of primers may be challenging due to the complex reaction sequence. To simplify the design process the use of a primer design software is recommended.
As sensitivity and non-template amplification of in-silico designed primers may vary, the evaluation of 2 - 4 real primer sets before choosing a final set is recommended.

Assay set-up
Pipet with sterile filter tips and perform the set-up in an area separate from DNA preparation or analysis. No-template controls should be included in all amplifications.
First, prepare a 10x conc. primer pre-mix. Second, set-up the isothermal amplification assay:
component||stock conc.||final conc.||Volumne 1x 20 μl Assay
Primer Mix||10x||1x||2 μl
Template DNA||||

Produtos relacionados:

• Saphir Bst Turbo Polymerase, #PCR-390
• Saphir Bst Turbo GreenMaster, #PCR-393
• Saphir Bst Turbo GreenMaster high ROX, #PCR-394