IGEM:metu/2009/Notebook/wound dressing/2009/08/27

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Image: Wound dressing logo.jpg

27.08.2009

1. Isolations of 1,2,3,4,7,9,11,BB and storages of 1,BB were made.

2. We made digestions. The following amounts are added to microtubes;

             Mono digestions:
  5,          water          18.82 µl                   7 µl
              Buffer Tango    3 µl        Buffer        4 µl
              Spe1            3 µl        EcoR1         2 µl       
              DNA             5.18 µl                  27 µl
  6,          water          13.67 µl                   6 µl
              Buffer O        3 µl        Buffer        4 µl 
              Pst1            3 µl        Xba1          2 µl       
              DNA            10.33 µl                  28 µl
  Pure plasmid double digestion;(6,8)
  (6 is cut with EcorI and Spe1 and 8 is cut with Xba1 and Pst1 restriction enzymes)
               water         11 µl
               Buffer         2 µl
               Enzyme       1+1 µl       6-EcoR1+Spe1   8-Xba1+Pst1
               DNA            5 µl
  Electrophoresis order;
  1) leader/ 6 / control 6 / control 8 / 8 / leader

            Control of double digestions:
  6,          water          19.4 µl                   21.4 µl
              FD Buffer       3 µl                      3 µl
              Xba1+Pst1     1+1 µl                      0
              DNA             5.6   =30 µl              5.6 µl   =30 µl
  8,          water          18.9 µl                    20.9 µl
              FD Buffer       3 µl                      3 µl
              Xba1+Pst1     1+1 µl                      0
              DNA             6.1 µl  =30 µl            6.1 µl    =30 µl
 We used 0.8% agorose gell and 10 µl EtBr
 Electrophoresis order;
 1) leader / 6 /  6 / control 6 / control 8 / 8 / 8 / leader

3. We couldn't digest.


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