1. Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends Ligation of the blunt end in pBR322A Selection of recombinant E. coli cells Isolation and purifictation of the pBR322 plasmid without BamH1 restrictionsite Digestion of pBR322  BamH1 with BamH1 and EcoRI restrictionenzymes Quality control by electrophoreses

2. Digestion of pBR322 with BamH1

3. Elongation of BamH1 restrictionsites, making of blunt ends

4. Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends

5. Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends Ligation of the blunt end in pBR322

6. Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends Ligation of the blunt end in pBR322A

7. Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends Ligation of the blunt end in pBR322A Selection of recombinant E. coli cells

8. Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends Ligation of the blunt end in pBR322A Selection of recombinant E. coli cells Isolation and purifictation of the pBR322 plasmid without BamH1 restrictionsite

9. Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends Ligation of the blunt end in pBR322A Selection of recombinant E. coli cells Isolation and purifictation of the pBR322 plasmid without BamH1 restrictionsite Digestion of pBR322  BamH1 with BamH1 and EcoRI restrictionenzymes

10. Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends Ligation of the blunt end in pBR322A Selection of recombinant E. coli cells Isolation and purifictation of the pBR322 plasmid without BamH1 restrictionsite Digestion of pBR322  BamH1 with BamH1 and EcoRI restrictionenzymes Quality control by electrophoreses

11. Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322  BamH1 plasmid Digestion of pBR322 with BamH1 Elongation of BamH1 restrictionsites, making of blunt ends Ligation of the blunt end in pBR322A Selection of recombinant E. coli cells Isolation and purifictation of the pBR322 plasmid without BamH1 restrictionsite Digestion of pBR322  BamH1 with BamH1 and EcoRI restrictionenzymes Quality control by electrophoreses