Thermo Fisher, K451020, TOPO™ TA Cloning™ Kit for Subcloning, with One Shot™ Mach1™ T1 Phage-Resistant Chemically Competent E. coli

TOPO™ TA Cloning™ Kit for Subcloning, with One Shot™ Mach1™ T1 Phage-Resistant Chemically Competent E. coli TOPO™ TA Cloning™ Kits for Subcloning provide a highly efficient, 5-minute, one-step cloning strategy ('TOPO™ cloning') for the direct insertion ofTaqpolymerase–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 cloning kits are available with a variety of competent cells, or without competent cells, depending upon your needs and budget. Features of the TOPO™ TA Cloning™ Kits for Subcloning:
•Fast and easy—go from PCR to clones in just 3 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 requiredTOPO™ TA Cloning™ Kits for Subcloning—overview
•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′-A overhangs (Taqpolymerase 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 ownpCR™ 2.
1-TOPO™ TA vector—cloning and subcloning simplicityThe pCR™ 2.
1-TOPO™ TA vector is linearized with 3'-thymidine (T) overhangs for direct ligation ofTaqpolymerase–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 manipulationThe pCR™ 2.
1-TOPO™ TA vector contains both ampicillin and kanamycin resistance markers and theLacZα gene for blue/white screening.
Simplified TOPO™ -based cloningUsing 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 transformE. colicompetent cells.
Efficient cloningWith up to 95% of clones carrying the desired insert, you can screen less clones, saving time and money. The pCR™ 2.
1-TOPO™ TA vector used in this kit comes with 3'-T overhangs for efficient ligation ofTaqpolymerase–amplified PCR products, which contain 3'-A overhangs.
The standard in cloningWhen 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 optionsThe 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. No. K4500-01, K4500-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4560-01, K4560-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. No. K4520-01, K4520-40)
• Fast growth: Mach1™ -T1R Chemically CompetentE. Coli(Cat. No. K4510-20)
• Provide your own: for flexibility and to save money (Cat. No. 450641)We also offer two versions of the kit that include a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4500-02 and K4510-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.
Bacterial or Yeast Strain: Mach1™
Cell Type: Chemically Competent
Cloning Method: TOPO™-TA
For Use With (Application): Chromatin Biology
Product Line: One Shot™
Product Type: Cloning Kit
Quantity: 25 Reactions
Vector: TOPO-TA Cloning Vectors
Format: Kit
Unit Size: Each