Team:UC Chile/Cyano/Notepad/week9

April 30 - May 6

Week 9

Monday 04.30

Did a massive PCR run. We amplified every part of Construct 1 using as template old PCR purifications and PCR products. We'll build C1 from scratch. Hoping everything works out well this time.

As an inspirational motif:

Muchos años después, al frente del pelotón de fusilamiento, el coronel Aureliano Buendía había de recordar aquella remota tarde en que su padre lo llevó a conocer el hielo.

Tuesday 05.01

We decided to transform e. coli with the expression plasmid pPMQAK1. Also, as the mentioned plasmid contains ccdb toxin, we took it out by a digestion/ligation procedure and transformed again. Transformed e. coli with GFP coding device.

During the afternoon, we checked yesterday's PCR's under UV light. Luckily, all parts amplified except for KanR u.u Did a PCR run for KanR alone. We tried with different master mix combinations, some containing GC buffer and some with DMSO.

Existential Question: WHAT DO YOU DO WHEN YOU HAVE A GREMLIN PLAGUE IN THE LABORATORY??

Got it!! The next protocol can be used,

Protocol to get rid of gremlins: http://christalawler.com/2010/06/05/bits-how-to-kill-a-gremlin-as-seen-in-gremlins/

• The previous protocol needs to be adapted to laboratory conditions!!!

Wednesday 05.02

Talked with one of our advisors today. He gave us interesting input on our failed PCR's.

Checked PCR runs. KanR didn't have the strenght we'd have liked, but still we used it. All parts were assembled by Gibson. Also, every part has its neg. control. After Gibson, we transformed coli in kan, chlor and both resistance plates.

gel image

Friday 05.04

Happy Star Wars day!!!! May the fourth be with you :)