Cosmid prep or scaled up mini prep

1. Grow 3-4 tubes of each cosmid in 5 ml media overnight
   
   
2. Harvest a total of 4 ml of each tube separately in 2 ml centrifuge tubes
   
   
3. Resuspend completely in 200 l Solution I*
   
   
4. Add 400 l Solution II* and invert until solution clears
   
   
5. Add 300 l Solution III* and invert gently
   
   
6. Hold on ice for 10 min
   
   
7. Spin 10 min at 4°C and transfer s/n to new tube
   
   
8. Phenol/chloro/IAA one time with equal volume
   
   
9. Spin 1 min at 4°C and transfer upper layer to new tube
   
   
10. Ppt with 0.8 vol isopropanol
    
    
11. Spin 20 min at 4°C
    
    
12. Remove s/n and allow pellet to dry
    
    
13. Resuspend in ~ 50 l of TE + RNase
    
    
14. Cosmids => Digest 5-10 l with either EcoRI or Hind III
    
    *refer to Alkaline Lysis Solution I, II, or III recipes in card box