pCR™8/GW/TOPO™ TA Cloning Kit with One Shot™ Mach1™-T1R E. coli
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Invitrogen™

pCR™8/GW/TOPO™ TA Cloning Kit with One Shot™ Mach1™-T1R E. coli

The pCR™8/GW/TOPO™ TA Cloning Kit delivers fast, efficient and reliable cloning. Additionally, this vector is ideal for use in Gateway™Read more
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Catalog number K252020
Price (USD)
626.00
Each
Add to cart
Price (USD)
626.00
Each
Add to cart
The pCR™8/GW/TOPO™ TA Cloning Kit delivers fast, efficient and reliable cloning. Additionally, this vector is ideal for use in Gateway™ recombination cloning into various destination vectors for a multitude of applications.

This pCR™8 cloning kit offers the following features:
• Fast, 5-minute TOPO™ TA Cloning delivers 95% recombinants
• 3´-T overhangs simplify cloning of Taq-amplified PCR products
• Fast growing MachI cells shorten cloning time
• Compatible with Gateway™ recombination cloning enabling access to a multitude of destination vectors
• Sequencing primer sites located close to cloning insertion site so you sequence more insert, less vector the first time
• Spectinomycin resistance gene for robust selection in E. coli
• EcoR I sites flanking the PCR product insertion site for easy excision of inserts

This kit includes One Shot™ Mach1™-T1R Chemically Competent E. coli.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Cloning MethodTOPO™ with Gateway™
No. of Reactions20 reactions
Product LineOne Shot™
Product TypeTA Cloning Kit
VectorpCR
Unit SizeEach
Contents & Storage
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, Gateway Forward and Reverse Sequencing Primers, Control Template and PCR Primers, 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

Frequently asked questions (FAQs)

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.

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