pFlag-CMV-TOPO Vector (20 rxns)

pFLAG-CMV-TOPO Vector is protein expression vector which has CMV promoter lead expression gene linked to FLAG peptide at N-terminal and has ampicillin antibiotic marker. It is transient mammalian expression vector which can produce high concentrate protein (0.1mg/L ~ 1mg/L). pFLAG-CMV-TOPO Vector is treated with Topoisomerase (topoisomerase is conjugated to each end of vector). So, ligation reaction is performed without ligase in a short time and high yield of cloning result can be expected.

Applications

- Cloning and expression of Taq polymeraseamplified genes in mammalian cells

- Detection of gene expression using anti-Flag antibody

Contents

- pFlag-CMV-TOPO Vector

- Salt Solution

- CMV-F Primer

- CMV-24R Primer

Standard reaction procedure

∙ PCR reaction

∙ Mix pFlag-CMV-TOPO Vector with PCR products

• PCR Product0.5~4 ㎕
  
• Salt Solution 1 ㎕
  
• Sterile Water up to 5 ㎕
  
• pFlag-CMV-TOPO Vector 1 ㎕
  
• Final Volume 6 ㎕
  

∙ Incubation at room temperature for 5min

∙ Transformation into E.coli

∙ Colony analysis

∙ Analysis of Cloned gene: orientation, open reading frame and sequence