Invitrogen™ TOPO™ TA Cloning™ Kit for Subcloning, with One Shot™ TOP10 chemically competent E. coli cells

Provides a highly efficient, 5-minute, one-step cloning strategy (“TOPO Cloning”) for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for subcloning

£547.20 valid until 31/12/2016
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Brand: Invitrogen™

Manufacturer Part Number: K450040


UNSPSC: 41106610

Code: 50

Additional Details:
Additional Details: Weight: 0.16000kg

Product Code. 10328252

Quantity Price
1 £760.00 / Each
EU Stock 81
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For Research Use Only. All usage must comply with product instructions.

Description and Specification


Product Type Kit, Subcloning
Bacterial/Yeast Strain TOP10
Includes Box 1: Topoisomerase I-activated pCR2.1-TOPO vector, PCR buffer, Salt solution, dNTPs, Control template, M13 forward and reverse primers, Control PCR primers, Sterile water. Box 2: One Shot Chemically Competent or Electrocomp E. coli, S.O.C. medium, Supercoiled pUC19 control plasmid
For Use With (Application) Cloning, PCR Cloning
Quantity 50 reactions

Cloning kits are available with a variety of competent cells, or without competent cells, depending upon your needs and budget. Each kit uses the pCR™ 2.1-TOPO TA vector with convenient restriction sites for subcloning.

pCR 2.1-TOPO TA vector: cloning and subcloning simplicity
The pCR 2.1-TOPO TA vector is linearized with 3'-thymidine (T) overhangs for direct ligation of Taq polymerase-amplified PCR products (TA cloning™) and is “activated” with covalently bound Topoisomerase I. There is no need to add ligase and cloning is complete in 5 minutes. EcoRI sites flanking the PCR product insertion site allow for easy excision of inserts or use any combination of 15 convenient restriction sites flanking your PCR insert for easy, directional subcloning.

pCR 2.1-TOPO TA clone selection and manipulation
The pCR 2.1-TOPO TA vector contains both ampicillin and kanamycin resistance markers and the LacZα gene for blue/white screening.

Simplified TOPO -based cloning
Using TOPO cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform E. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen fewer clones, saving time and money. The pCR 2.1-TOPO TA vector used in this kit comes with 3'-T overhangs for efficient ligation of Taq polymerase-amplified PCR products, which contain 3'-A overhangs.

The standard in cloning
When it comes to cloning, TOPO™ cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO cloning has been applied to many different vectors for a wide array of applications.

TOPO TA Cloning Kits for Subcloning: kit options
The TOPO TA Cloning Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

• General cloning: TOP10 cells (Cat. Nos. K4500-01, K4500-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. Nos. K4560-01, K4560-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. Nos. K4520-01, K4520-40)
• Fast growth: Mach1™ -T1R Chemically Competent E. Coli (Cat. No. K4510-20)
• Provide your own: for flexibility and to save money (Cat. No. 450641)

Two versions of the kit that include a PureLink™ Quick Plasmid Miniprep Kit (Cat. Nos. K4500-02 and K4510-02) for use in isolation of clean, sequencing-ready, recombinant plasmid are also available.

• Fast and easy: go from PCR to clones in just three steps and in as little as 5 minutes hands-on time
• Efficient: obtain up to 95% clones with correct insert
• Proven: reliable performance for over a decade with over 4,000 citations
• Simple: no ligase, post-PCR procedures, or PCR primers containing specific sequences are required
• Vector: pCR 2.1-TOPO TA vector: subcloning vector with 15 convenient restriction sites flanking your insert for easy, directional subcloning
• Cloning method: TOPO TA Cloning : Topoisomerase I-based, 5-minute ligation of PCR products with 3 ft.-A overhangs (Taq polymerase amplified) to TOPO vector with T overhangs
• Competent cells: various options: choose from kits with either general, high-efficiency, bacteriophage T1-resistant, fast-growing competent cells, or use your own


Cloning, PCR Cloning

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