Product Description
TOP10F ́, Linearized pCRII vector, ExpressLink T4 DNA ligase, 5X ExpressLink T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, controls, One Shot TOP10F' chemically competent E. coli, S.
O.
C. medium, and a supercoiled control plasmid., 40 Reactions The TA Cloning™ Kit Dual Promoter with pCR™II vector provides a quick, one-step cloning strategy for directly inserting aTaq-amplified PCR product into a plasmid vector. The T7 and Sp6 promoters of the pCR™II vector allowin vitrotranscription of the insert to produce sense or anti-sense products. The TA Cloning Kit Dual Promoter uses the pCR™II cloning vector and ExpressLink™ T4 DNA Ligase to generate a ligation product in a fifteen-minute, room-temperature ligation step. Reactions typically yield >80% recombinants containing inserts.
Features of the TA Cloning™ Kit Dual Promoter:
•Fast & convenient—15-minute, room-temperature ligation
•Efficient—blue/white screening and >80% clones with correct insert
•Flexible—choice of kanamycin or ampicillin resistance for flexible antibiotic selection
•Hassle-free—eliminates any enzymatic modifications of the PCR product
•Streamlined—does not require the use of PCR primers that contain restriction sitesThe pCR™II vector provides:
• 3'-T overhangs for direct ligation ofTaq-amplified PCR products
• T7 and Sp6 promoters forin vitroRNA transcription and sequencing
• Versatile polylinker with flankingEcoR I sites for easy excision of inserts
• M13 forward and reverse primer sites for sequencingHow TA Cloning™ WorksTaqpolymerase has a non-template-dependent activity that adds a single deoxyadenosine (A) to the 3' ends of PCR products. The linearized vector supplied in this kit has single 3' deoxythymidine (T) residues. This allows PCR inserts to ligate efficiently with the vector.
Kit ConfigurationsThe TA Cloning™ Kit is offered in a variety of configurations: with One Shot™ INVF' Chemically CompetentE. coli(K2050-01 and K2050-40), One Shot™ TOP10F' Chemically CompetentE. coli(K2060-01 and K2060-40), and without competent cells (K2750-20 and K2750-40) in 20- and 40-reaction kit sizes.
Bacterial Content: TOP10
Bacterial or Yeast Strain: TOP10F ́
Cell Type: Chemically Competent E. coli
Cloning Method: TA Cloning
For Use With (Application): PCR Cloning
Includes: Linearized pCRII vector, ExpressLink T4 DNA ligase, 5X ExpressLink T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, controls, One Shot TOP10F' chemically competentE. coli, S.
O.
C. medium, and a supercoiled control plasmid.
No. of Reactions: 40 Reactions
Product Line: One Shot™
Product Type: Cloning Kit
Promoter: T7, SP6
Quantity: 40 rxns
Vector: pCRII
Format: Kit
Unit Size: Each
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Collaboration
Tony Tang
Email: Tony.Tang@iright.com
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