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

From OpenWetWare
Jump to: navigation, search
Igem-logo-150px.png WOUND DRESSING Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

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.