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

Catalog number: K4500J10

Invitrogen™  Related applications: PCR Cloning

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Description

TOPO® TA Cloning® kits for subcloning provide a highly efficient, 5-minute cloning reaction for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for subcloning. Each kit uses the pCR™2.1-TOPO® TA vector with convenient restriction sites for subcloning. The TOPO® TA Cloning® kits are available with a variety of competent cells, or without competent cells, depending upon your needs and budget.

• Fast and easy—go from PCR to clones in just 3 steps, including a 5-minute cloning reaction
• Efficient—obtain up to 95% clones with correct insert
• Proven—enabling reliable performance for over a decade with over 20,000 citations
• Simple—no ligase, post-PCR procedures, or PCR primers containing specific sequences required

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 (which have 3'-A overhangs) and "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.

The Most Widely Used Cloning Kit
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.

Kit Options: TOPO® TA Cloning® Kits for Subcloning
TOPO® TA Cloning® kits for subcloning can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:
• General cloning: TOP10 cells (Cat. Nos. K4500-J10, 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 cells (Cat. Nos. 451641, 450641)

We also offer 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.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Vector Type: TOPO-TA Vectors
Cloning Method: TOPO®-TA
Bacterial or Yeast Strain: TOP10
Antibiotic Resistance (Bacterial): Ampicillin (AmpR), Kanamycin (KanR)
Product Size: 10 reactions

Contents & storage

Box 1:
• Topoisomerase I-activated pCR™2.1-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.

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