I gotta feeling!!!
- After inactivating the restriction, I ran a gel to check if everything went on well. Though the gel merely shows a band, I consider ok to follow on with the ligation.
- The lanes were as follows:
1, 5. Ladder.
2. Click Beetle Green luciferase restricted with XBA I and PST I.
3. Click Beetle Red luciferase restricted with XBA I and PST I.
- The next reactions describe the ligations to be done for a total of 20μl.
- 1: CBG (XBA I/PST I) with J23101 labeled as CBG + J23101 Mar and date.
-H2O -----------------------> 7μL
-Buffer ligase -----------> 4μL
-Vector (plasmid) -----> 2μL
-Insert (CBG) ---> 6μL
-Ligase --------------------> 1μL
- 2: CBR (XBA I/PST I) with J23101 labeled as CBR + J23101 Mar and date.
-H2O -----------------------> 7μL
-Buffer ligase -----------> 4μL
-Vector (plasmid) -----> 2μL
-Insert (CBR) ---> 6μL
-Ligase --------------------> 1μL
- I also replated 5 strains of pT7 + luciferase per transformation into kanamycin plates.
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