Invitrogen™ TA Cloning™ Kit, Dual Promoter, with pCR™II Vector and One Shot™ INVαF' Chemically Competent E. coli

Provides quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into plasmid vector

Product Code	Brand	Bacterial or Yeast Strain	Includes	No. of Reactions
10003102	Invitrogen™ K205001	INVαF ́	Linearized pCRII vector, ExpressLink T4 DNA ligase, 5X ExpressLink T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, controls, One Shot INVαF' chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid.	20 Reactions
10740165	Invitrogen™ K205040	INVαF ́	Linearized pCRII vector, ExpressLink T4 DNA ligase, 5X ExpressLink T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, controls, One Shot INVαF' chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid.	40 Reactions

Description

The T7 and Sp6 promoters of the pCRII vector allow in vitro transcription of the insert to produce sense or anti-sense products. The TA Cloning Kit Dual Promoter uses the pCRII 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.

 

• Fast and 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 PCR product
• Streamlined: does not require use of PCR primers that contain restriction sites
• 3'-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and Sp6 promoters for in vitro RNA transcription and sequencing
• Versatile polylinker with flanking EcoR I sites for easy excision of inserts
• M13 forward and reverse primer sites for sequencing

 

 

• How TA Cloning Works\
• Taq polymerase has non-template-dependent activity that adds single deoxyadenosine (A) to 3' ends of PCR products
• Linearized vector supplied in this kit has single 3' deoxythymidine (T) residues
• Allows PCR inserts to ligate efficiently with vector
• Kit Configurations
• TA Cloning Kit is offered in a variety of configurations: with One Shot INVF' Chemically Competent E. coli (K2050-01 and K2050-40), One Shot TOP10F' Chemically Competent E. coli (K2060-01 and K2060-40), and without competent cells (K2750-20 and K2750-40) in 20- and 40-reaction kit sizes

 

Cloning, PCR Cloning

Specifications

• Bacterial or Yeast Strain: INVαF ́
• Cloning Method: TA Cloning
• Format: Kit
• Includes: Linearized pCRII vector, ExpressLink T4 DNA ligase, 5X ExpressLink T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, controls, One Shot INVαF' chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid.
• Product Type: Cloning Kit
• Vector: pCRII

• Cell Type: Chemically Competent E. coli
• Content And Storage: This kit contains linearized pCR™II vector, ExpressLink™ T4 DNA ligase, 5X ExpressLink™ T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, and controls. Competent cell kits contain One Shot™ chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid.
  
  Store One Shot™ E. coli at -80°C. Store all other components at -20°C. All reagents are guaranteed stable for 6 months when properly stored.
• For Use With (Application): PCR Cloning
• Product Line: One Shot™
• Promoter: T7, SP6

Safety and Handling

Product Identifier

• TA Cloning Kit, Dual Promoter, with pCRII Vector and One Shot INV

Signal Word

• Warning

Hazard Category

• Reproductive toxicity Category 2
• Skin sensitiser Category 1

Hazard Statement

• H317-May cause an allergic skin reaction.
• H361-Suspected of damaging fertility or the unborn child.

Precautionary Statement

• P201-Obtain special instructions before use.
• P202-Do not handle until all safety precautions have been read and understood.
• P261-Avoid breathing dust/fume/gas/mist/vapours/spray.
• P272-Contaminated work clothing should not be allowed out of the workplace.
• P273-Avoid release to the environment.
• P280-Wear protective gloves/protective clothing/eye protection/face protection.
• P302+P352-IF ON SKIN: Wash with plenty of water/soap.
• P308+P313-IF exposed or concerned: Get medical advice/attention.
• P333+P313-If skin irritation or rash occurs: Get medical advice/attention.
• P362+P364-Take off contaminated clothing and wash it before reuse.
• P501b-Dispose of contents/container in accordance with local/regional/national/international regulations.

Supplemental information

• MIXTURE LIST-Contains : 2-Mercaptoethanol, Hydrogen chloride

For Research Use Only. Not for use in diagnostic procedures.