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Team:Colombia Uniandes/NickoJournal
From 2013.igem.org
Contents |
Dear Journal! :)
Here we will place all the information about the work we have been doing during this time, it will be named by date. We hope you enjoy our work and thoughts as much as we do!
18-Jun-2013
Hello! Hello! Hola! Hola! After we went over the project, we proceed to design our primers so we could place our order, taking into account that we are going to use Fusion PCR as our main protocol. Our primers take around 2 weeks to arrive… sooo meanwhilee all the team started making our E.coli babies electrocompetent, so we can work with them later on.
25-Jun-2013
So today we took our bacterial cells (Top 10) onto an LB medium (no antibiotics). We let it ON at 37°C on the shaker. Also we prepared all the material we need for tomorrow: ddwater, 10% glycerol, LB medium and microfuge tubes.
26-Jun-2013
Today we are going to start preparing our cells, we inoculated in the morning and placed all our material that needs to be cold in the fridge 4°C(ddWater and glycerol), also the centifuge rotor needs to be chilled. Then we just waited for the perfect OD and made our cells around 4:00pm. We made a lot!
We placed them in the -80, ready to start our work!!!
2-Jul-2013
Our primers are FINALLY here and we are anxious to start working!!! So we made our plan(In theory)on big steps for the next weeks!:
1. DNA extraction from E.coli and Ralstonia metallidurans
2. PCR
2.1 PrcnR, RBS, rcnR, stop (Amplified as one piece) 2.2 PrcnA,RBS 2.3 hoxN, stop
3. Fusion PCR
3.1 PrcnA/hoxN 3.2 PrcnR,RBS,rcnR/PrcnA/hoxN
On each PCR add 30 steps more of making gels for confirmation, again and again and again.
3-Jul-2013
Fortunately our team member Silvia Cañas already had a DNA extraction from E.coli so she donate it to us so we could start working! Thank you Silvia! :)
We asked for Ralstonia metallidurans to our teacher Jenny Dussan at her lab here in the university. She is giving us the strain tomorrow morning! Thank you Jenny!
This done, did the extraction of R.metallidurans and store it in the -30°C.
