Maxiprep   to Extract Plasmids

 

Materials:

Methods:

1. Follow Protocol Appendix B.2 for QIAGIN-tip 500 (on 100 ml of high copy number plasmid). Redissolve the DNA in 0.5    10 mM Tris-HCl (pH 8.5).
2. Estimate DNA concentration using spectrophotometry: after blanking with 700    10 mM Tris-HCl cuvette, aliquot 50   from DNA solution into 600   10 mM Tris-HCl cuvette.
3. Run gel electorphoresis by Following protocol in Appendix E.1, with the following lanes:

   

Results: ( 2/7/99)

• For DNA concentration: for 50  /600ml.

  dilution factor = 12:1

   /ml = 6.365  /ml

  6.365  /ml (dilution factor)

• Problem with gel loading: maxiprep debug samples did not settle in the gel, probably because the Tris-HCl buffer was too light. Should have used only 2   from DNA-debug/Tris-HCL solution, and rest   as buffer. Therefore, have a 3 lane separation between maxiprep debug samples, and ladder and pGFP(LVA) samples. Figure 2.1(b) shows the gel, with pGFP(LVA) showing nicely in the right two lanes.