Master mix Taq Pol (2X) Green

Master mix para carregamento direto em géis de eletroforese

Pronto para uso, verde

Catálogo Nº Apresentação Preço (R$) Comprar
POL-102XS 50 reações130,00 Adicionar ao Carrinho
POL-102S 100 reações260,00 Adicionar ao Carrinho
POL-102M 200 reações400,00 Adicionar ao Carrinho
POL-102L 500 reações900,00 Adicionar ao Carrinho
POL-102XL 1.000 reações1.250,00 Adicionar ao Carrinho

For in vitro use only!

Envio: Shipped on blue ice

Condições de armazenamento: Store at -20 °C
Avoid freeze/thaw cycles. Taq Pol – Master mix (2X) is also stable for three months at 4°C, so for frequent use, an aliquot may be kept at 4°C

Validade: 12 months

Forma: Liquid

Concentração: 2x concentrated

Descrição:
Taq Pol Master Mix Geen contains Taq Polymerase in an optimized PCR buffer with Mg2+ and dNTPs. It contains all reagents required for PCR (except template and primer) in a premixed 2x concentrated ready-to-use solution and should be used at a 1X concentration with DNA template and primers in a total reaction volume of 25 or 50 μL. The Master Mix is recommended for use in routine PCR reactions from templates including pure DNA solutions, bacterial colonies, and cDNA. It is optimized for high specificity and guarantees minimal by-product formation. The tracking dyes in the master mix do not interfere with PCR performance and are compatible with downstream applications such as fluorescent automatic DNA sequencing, ligation, and restriction digestion. The blue dye migrates with 3-5 kb fragments and the yellow dye migrates faster than 20 bp (1% agarose gel).

Contente:
2x Taq Pol Master Mix Green (violet cap)
Composition: 0.05 U/ μL Taq DNA polymerase, reaction buffer, 0.3 mM MgCl2, 0.4 mM of each dNTP (dATP, dCTP, dGTP, dTTP), dye, gel loading buffer and stabilizers. and stabilizers.


PCR Reaction Setup:
Use the quantities below to prepare a single 50 μl PCR reaction. Thaw, mix, and briefly centrifuge each component before use.

Add the following components to a microcentrifuge tube:

1. Prepare PCR master mix
Note: Consider the volumes for all components listed next steps to determine the correct amount of water required to reach your final reaction volume.

component50 μl
assay
final conc.
PCR grade waterto 50 μl-
2x Taq Pol Master Mix25 μl1x


Mix and briefly centrifuge the components.
2. Add template DNA and primers

Component50 μl
assay
final conc.
Forward primer (10 μM) 0,5 – 2,5 μl 0,1 – 0,5 μM
Reverse primer (10 μM) 0,5 – 2,5 μl 0,1 – 0,5 μM
Template DNAx μl10 pg – 1 μg*


*genomic DNA: 1 ng – 1 μg; plasmidial DNA: 1 pg – 1 ng
Cap each tube, mix, and briefly centrifuge the content.
4. Incubate reactions in a thermal cycler
Recommended cycling conditions:

Initial denaturation 95 °C1 – 3 min1x
Denaturation95 °C15 - 30 sec30 cycles
Annealing**45 - 68 °C 15 - 30 sec30 cycles
Elongation***72 °C 1 min/kb30 cycles
Final extension (optional) 72 °C 2 min/kb1x
Hold4 – 8 °C 1x


**The annealing temperature depends on the melting temperature of the primers used
***The elongation time depends on the length of the fragments to be amplified. A time of 1 min/kb is recommended.
For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.