Invitrogen™ pcDNA™3.3-TOPO™ TA Cloning™ Kit

Invitrogen™ pcDNA™3.3-TOPO™ TA Cloning™ Kit

The pcDNA™3.3-TOPO® TA Cloning® Kit is designed to deliver exceptionally high levels of transgene expression in adherent cells as well as in the FreeStyle™ MAX CHO and FreeStyle™ MAX 293 suspension cell systems. The pcDNA3.3™-TOPO® TA Cloning® Kit is supplied with the TOPO®-adapted plasmid vector, all reagents for cloning, and One Shot® TOP10 competent cells.

Features

• TOPO® adapted for cloning PCR products with >85% efficiency
• High-level gene expression in a wide range of mammalian cells due to the native cytomegalovirus (CMV) immediate-early promoter/enhancer sequence (672 bp)
• Vector is ideal for large-scale protein expression
• Generation of native protein without extraneous amino acids

Contents

Box 1:

• pcDNA™3.3-TOPO™ vector, TOPO™ adapted (5–10 ng/µl): 20 µl 
• 10X PCR Buffer: 100 µl 
• dNTP Mix: 10 µl 
• Salt Solution: 50 µl 
• Sterile Water: 1 ml 
• Control PCR Template (50 ng/µl): 10 µl
• Control PCR Primers (100 ng/µl each): 10 µl 
• CMV Forward Sequencing Primer (100 ng/µl): 20 µl 
• TK polyA Reverse Sequencing Primer (100 ng/µl): 20 µl 
• pcDNA™3.3-TOPO™/lacZ Expression Control Plasmid (500 ng/µl): 10 µl 

Box 2 

• TOP10 E. coli: 11 × 50 µl
• pUC19 Control DNA (10 pg/µl): 50 µl 
• S.O.C. Medium: • 6 ml