TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells
TOPO&trade; TA Cloning&trade; Kit, Dual Promoter, with One Shot&trade; TOP10 chemically competent <i>E. coli</i> cells
Invitrogen™

TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells

TOPO™ TA Cloning™ Dual Promoter kits are for fast, efficient cloning and subsequent in vitro transcription. These kits include theRead more
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Catalog NumberQuantity
K46000125 Reactions
K4600J1010 Reactions
K46004050 Reactions
Catalog number K460001
Price (USD)
746.00
Each
Add to cart
Quantity:
25 Reactions
Price (USD)
746.00
Each
Add to cart
TOPO™ TA Cloning™ Dual Promoter kits are for fast, efficient cloning and subsequent in vitro transcription. These kits include the pCR™II-TOPO™ TA vector with dual T7 and SP6 promoters. By eliminating time-consuming and tedious restriction site cloning, TOPO™ cloning is the most reliable cloning method, featuring a 3-step protocol and 5-minute cloning reaction, and yielding up to 95% recombinants.

Convenient features of this TOPO™ TA Cloning™ Dual Promoter kit include:
• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and SP6 promoters for efficient in vitro transcription
• M13 forward and reverse primer sites for sequencing
• 16 convenient restriction sites, including EcoRI, flanking your insert for subsequent excision or subcloning
• Kanamycin and ampicillin resistance for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants
• Includes OneShot™ TOP10 Competent cells for added convenience and highest cloning efficiencies

Kit Options: TOPO™ TA Cloning™ Dual Promoter Kits
TOPO™ TA Cloning™ Dual Promoter kits can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:
• General cloning: TOP10 cells (Cat. Nos. K4600-J10, K4600-01, K4600-40)
• High-efficiency cloning: TOP10 Electrocomp™ cells (Cat. Nos. K4660-01, K4660-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. Nos. K4620-01, K4620-40)
• Fast growth: Mach1™ -T1R chemically competent E. coli (Cat. No. K4610-20)
• Repressor/induction needs: TOP10F’ cells (Cat. Nos. K4650-01, K4650-40)
• Provide your own cells (Cat. Nos. 451641, 450641)
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Bacterial or Yeast StrainTOP10
Cell TypeChemically Competent
Cloning MethodTOPO™-TA
FormatKit
Product LineOne Shot™
Product TypeCloning Kit
PromoterT7, SP6
Quantity25 Reactions
VectorTOPO-TA Cloning Vectors
Unit SizeEach
Contents & Storage
Box 1:
• Topoisomerase I-activated pCR™II-TOPO™ vector
• PCR buffer
• Salt solution
• dNTPs
• Control template
• M13 forward and reverse primers
• Control PCR primers
• Sterile water

Store at -5 to -30°C.
All reagents are stable for 6 months when properly stored.

Box 2:
• One Shot™ Chemically Competent or Electrocomp™ E. coli
• S.O.C. medium
• Supercoiled pUC19 control plasmid.

Store at -68 to -85°C.

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.

What is the difference between the pCR2.1-TOPO and pCR4-TOPO vectors?

The vector backbones for both of these vectors are very similar. The main difference is that the pCR4-TOPO vector has sequencing primer sites located as close as 33 base pairs from the PCR product insertion site. This minimizes the amount of vector DNA sequence that needs to be read before reaching the sequence of the insert, making the pCR4-TOPO vector very useful for sequencing applications.

What is the difference between the pCR2.1-TOPO and pCRII-TOPO vectors?

The vector backbones for both of these vectors are very similar. The main difference is that the pCRII-TOPO vector is a dual promoter vector, containing the SP6 and T7 promoters for in vitro transcription/sequencing, whereas the pCR2.1-TOPO vector contains only the T7 promoter for in vitro transcription/sequencing. Both vectors contain the M13 Forward and Reverse primer sites for sequencing or PCR screening.

Your TOPO cloning kits contain a control template and control primers. Can I obtain the sequence of the control template?

The sequence of the control template is proprietary.