Time is running out!
- Today, I extracted plasmid from the green click bettle luciferase culture and I left a restriction for 20μl to see if the plasmid contains the luciferase. I also made a restriction with SPEI and PST I to be able to ligate it with LRE restricted with XBAI and PST I.
- I also made some plates with Cm 30 and Am 30 to do the cotransformation.
- In the afternoon, I ran a gel to check if the restriction went ok, the gel is shown next:
- The lanes were as follows:
2. Restricted GCB luciferase with ECO RI and PST I.
3. Restricted GCB luciferase with SPEI and PST I.
4. Restricted LRE with XBAI and PST I.
- Unfortunately, I keep observing these awful dots in the CB luciferases, that is why I ran a gel of both, the plasmid purification and the respective restriction with ECO RI and PST I to try to infer what is happening, the gel is shown next:
- The lanes from right to left are as follows:
1, 2. Ladder
3, 5. Plasmid restriction
4, 6. Plasmid purification