Create pBR322+GFP(LVA)

 

Attempt to make unique ligation sites:

• Cut pBR322 (4361 bases) with ``AatII'' (at 4284) and ``EcoRI'' (at 4359) restriction enzymes. AatII has the following matching sequence:

  

  EcoRI has the following matching sequence:

  

• Cut pGFP(LVA) (3kb) with ``XbaI'' (at 274) and HindIII (at 1062) restriction enzymes. XbaI has the following matching sequence:

  

  HindIII has the following matching sequence:

  

 
• Digest pBR322 and pGFP(LVA) with 2 Enzymes Each
• Gel Purify Digested pBR322 and GFP(LVA)
• Anneal XbaI-EcoRI Adapter
• Ligate pBR322, GFP(LVA), adapters
• Gel Purify Ligation of pBR322 + GFP(LVA)