1. Preparation of competent E. coli cells Preparation of probes by PCR Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification of the digested fragment Ligation of fragments with pBR322 plasmid Transformation of the ligated construct into pBR322 Selection of recombinant E. coli cells Isolation and purification of the pBR322 plasmid with insert Digestion of pBR322 with restriction enzymes.Identification of inserts by electrophoreses and Southern blotting

2. Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV

3. Purification of the digested fragment

4. Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification of the digested fragment Ligation of fragments with pBR322 plasmid

5. Preparation of competent E. coli cells Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification of the digested fragment Ligation of fragments with pBR322 plasmid Transformation of the ligated construct into pBR322

6. Preparation of competent E. coli cells Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification of the digested fragment Ligation of fragments with pBR322 plasmid Transformation of the ligated construct into pBR322 Selection of recombinant E. coli cells

7. Preparation of competent E. coli cells Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification of the digested fragment Ligation of fragments with pBR322 plasmid Transformation of the ligated construct into pBR322 Selection of recombinant E. coli cells Isolation and purification of the pBR322 plasmid with insert

8. Preparation of competent E. coli cells Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification of the digested fragment Ligation of fragments with pBR322 plasmid Transformation of the ligated construct into pBR322 Selection of recombinant E. coli cells Isolation and purification of the pBR322 plasmid with insert Digestion of pBR322 with restriction enzymes

9. Preparation of competent E. coli cells Preparation of probes by PCR Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification of the digested fragment Ligation of fragments with pBR322 plasmid Transformation of the ligated construct into pBR322 Selection of recombinant E. coli cells Isolation and purification of the pBR322 plasmid with insert Digestion of pBR322 with restriction enzymes.Identification of inserts by electrophoreses and Southern blotting

10. Preparation of competent E. coli cells Preparation of probes by PCR Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification of the digested fragment Ligation of fragments with pBR322 plasmid Transformation of the ligated construct into pBR322 Selection of recombinant E. coli cells Isolation and purification of the pBR322 plasmid with insert Digestion of pBR322 with restriction enzymes.Identification of inserts by electrophoreses and Southern blotting