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

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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.