Saphir Bst Turbo GreenMaster

Master mix for isothermal DNA amplification with EvaGreen

Isothermal Amplification

Catálogo Nº Apresentação Preço (R$) Comprar
PCR-393S 2 x 1,25 mlSob demanda Adicionar ao Carrinho
PCR-393L 10 x 1,25 mlSob demanda Adicionar ao Carrinho

For general laboratory use.

Envio: shipped on gel packs

Condições de armazenamento: store at -20 °C
store dark
Short term storage (up to 3 months) at 4 °C possible.

Validade: 12 months

Forma: liquid

Concentração: 2x conc.

Propriedades espectroscópicas: λexc 500 nm (EvaGreen bound to DNA), λem 530 nm (EvaGreen bound to DNA)

Descrição:
Saphir Bst Turbo GreenMaster is a complete 2x conc. master mix 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
Saphier Bst Turbo Polymerase, dNTPs, reaction buffer, glycerol, EvaGreen DNA intercalator dye, stabilizers

PCR-grade water

Detection
The mix contains the fluorescent DNA stain EvaGreen® that intercalates into DNA during the amplification process and allows the direct quantification of target DNA by fluorescence detection (analogous to real-time PCR).
The mix can be combined with ROX reference dye (#PCR-351) to allow a signal normalization in real-time PCR instruments that are compatible with the evaluation of the ROX signal.

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 5-10 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
A reaction volume of 20-50 μl is recommended for most applications. 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.||20 μl||50 μl
Saphir Bst Turbo GreenMaster||2x||1x||10 μl||25 μl
Primer Mix||10x||1x||2 μl||5 μl
Template DNA||||

Produtos relacionados:

  • Saphir Bst Turbo Polymerase, #PCR-390
  • EvaGreen DNA Stain, #PCR-379
  • MgCl2 Solution, #PCR-266
  • dNTP Mix / 10 mM, #NU-1006
  • dNTP Mix / 25 mM, #NU-1023