pCR™8/GW/TOPO™ TA Cloning Kit with One Shot™ Mach1™-T1R Chemically Competent E. coli (Supply Center Only)
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Invitrogen™

pCR™8/GW/TOPO™ TA Cloning Kit with One Shot™ Mach1™-T1R Chemically Competent E. coli (Supply Center Only)

TOPO™ TA Cloning™ Kits are designed for cloning PCR products directly from a PCR reaction in just 5 minutes. TOPO™Más información
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Número de catálogoCantidad
K252020SC20 reacciones
Número de catálogo K252020SC
Precio (CLP)
751.138
Each
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Cantidad:
20 reacciones
Precio (CLP)
751.138
Each
Añadir al carro de la compra
TOPO™ TA Cloning™ Kits are designed for cloning PCR products directly from a PCR reaction in just 5 minutes. TOPO™ TA Cloning™ Kits use a pCR™-TOPO™ Vector with covalently bound topoisomerase I for fast cloning and recombinants. The newest TOPO™ TA Cloning™ Kit comes with a Gateway™ (GW) entry vector, pCR™8/GW/TOPO™ (Figure 1). This vector has:

• 3´-T overhangs for direct ligation of Taq-amplified PCR products
att sites for rapid recombination into a variety of Gateway™ destination vectors
• Novel GW primer sites, within the att regions, located less than 55 base pairs from the PCR product insertion site for convenient sequencing
• Spectinomycin resistance gene for robust selection in E. coli
EcoR I sites flanking the PCR product insertion site for easy excision of inserts

Selected TOPO™ TA Cloning™ Kits for entry into the Gateway™ Technology are available in combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO™-cloned inserts for downstream analysis.


Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones
Método de clonaciónTOPO™ with Gateway™
N.º de reacciones20 reactions
Línea de productosOne Shot
Tipo de productoTA Cloning Kit
Cantidad20 reacciones
Condiciones de envíoDry Ice
VectorpCR
Unit SizeEach
Contenido y almacenamiento
The pCR™8/GW/TOPO™ TA Cloning Kit contains the TOPO™-adapted, ready-to-use pCR™8/GW/TOPO™ Vector, dNTPs, Salt Solution, Sterile Water, GW Forward and Reverse Sequencing Primers, Control Template and PCR Primers, One Shot™ TOP10 Chemically Competent E. coli or One Shot™ Mach1™-T1R Chemically Competent E. coli, S.O.C. Medium, and a pUC19 control plasmid. Store the One Shot™ E. coli at -80°C. Store all other components at -20°C. All reagents are guaranteed stable for 6 months when properly stored.

Preguntas frecuentes

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

Your Gateway-adapted TOPO vectors are supplied with a control template and control primers. Can I obtain the sequence of the control template?

The sequence of the control template is proprietary.

What is the best ratio of insert:vector to use for cloning? Is there an equation to calculate this?

The optimal ratio is 1:1 insert to vector. Optimization can be done using a ratio of 0.5-2 molecules of insert for every molecule of the vector.

Equation:

length of insert (bp)/length of vector (bp) x ng of vector = ng of insert needed for 1:1 insert:vector ratio

Does Platinum Taq DNA Polymerase High Fidelity enzyme mix leave 3' A-overhangs on the PCR product for subsequent cloning into a TOPO TA or original TA vector?

Yes, the enzyme mix leaves 3' A-overhangs on a portion of the PCR products. However, the cloning efficiency is greatly decreased compared to that obtained with Taq polymerase alone. It is recommended to add 3' A-overhangs to the product for TA cloning.