Team:UC Chile/Cyano/Notepad/week17

June 25 - July 1

Monday: first day of pseudo-vacations! Classes are over and just the exams are remaining. Simon and Carla believe they don't have any left (the grades of the last test are not published yet), but they are confident. They are full time iGEM now.

Tuesday: Tried Gibson with new enzymes (again). KanR was amplified by PCR (finally after four failed attempts!!). Also, we invited Dr. Endy to our Synbio Workshop in December and he kindly said he'd come (yeah!).


Wednesday: Gibson did not work this time so now we believe the temperature is the cause (we keep the DNA in ice throughout the whole procedure until the heat shock). This reaction worked in summer. Winter has come and we are waaaaaaay beyond the wall. Tried Gibson at different room temperature exposure times before heat shock: no exposure time, 5 seconds, 15 seconds and room temperature.


Also, Simon and Carla found out they didn't skip the exams they thought they have so the mood is kind of low. And after the failed Gibson they decided to go drinking after the lab work is done.

Thursday:

Results of previous experiment:

- no exposure = +
- room temperature = +
- 5 sec = -
- 15 sec = -

Conclusion: We have extremists bacteria!!!

Well, did one more Gibson at room temperature... waiting for results.

Just got off the phone with Fernán (the Argentinian). He says: "Our Gibsons haven't worked lately, I think that the cells are lacking... The are only 10´9 colonies per ug of pUC19 DNA." 10´9? Thats just 10 to 50 more times than ours! ... Starting Knight's TOP10 Competent protocol again (3 time this year)!

A centrifuge just exploded in the lab next door. Simon is an hero (turned off the centrifuge).

Also, we received an integration plasmid from Utah State former iGEM team. Thanks guys!!!