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

WOUND DRESSING

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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.