Team:Poznan-BioInf/Book

Prologue.

October 2012. Weronika came back from the iGEM 2012 Regional Jamborees. After a series of Poznan University of Technology Bioinformatics Students' Research Group meetings concerning synthetic biology Poznan-BioInf came into being.

Cold, cold November. We've participated in "Generation of the Future" programme organized by the Ministry of Science and Higher Education for students needing financial support to take up a challenge in an international contest. And we're waiting... ...and waiting... and waaaaaiting... and at the end of April 2013 we've received decision that we are granted 200 000 zlotys.

May. we've officially signed the funding agreement and we're waiting for approval. Unfortunatelly two thirds of out team were working on end-of-term examination and Bachelor's thesis so... we received BioBricks and we're waiting for decision that we can administer our funds. That month We are looking for wet lab instructor and place, where we could spend summer. We are going to have meeting to one of our professor, who we are going to ask about help. Finally we where send to Phd Przemyslaw Nuc, who we welcomed in our team.

And then came June. We are still waiting for agreement acceptation. We've started to transform E. coli with biobirck form parts registry to test the efficiency of our strain and our protocols. Since now we are working on borrowed laboratory equipments. During the tests we're looking for a suitable GFP. From now on we start to working with BBa_E0040, which exhibits bright green fluorescence.

June.

1st-4th week

We've started to trying to equip our lab. Because we are studying on two universities: Poznan University of Technology (where we officially belong) and Adam Mickiewicz University, we had some problems with the administration. On Poznan University of Technology there is no wet laboratory specialized at microbiological research. That's why we are force to work on Adam Mickiewicz University.

Problems have started to rise. Due to Polish law we can't make order as fast as we think and as we would like to. Under all (every single one) of our orders we need four various signs. What is more, we are oblige to contact three various producers or distributor to get valuation of their products. Unfortunately to us the holidays begins so we encounter obstructs with making an appointments.

July.

5th-6th week

We shake off from the shock. We are still trying to make an order at the biotechnological companies, so.. we are waiting for the reply from distributors.

7th week

We are still trying to make an order at the biotechnological companies, so.. we are gathering the signatures and forcing the offices of administration.

8th week

We are still waiting...

August.

9th-10th week

We're done with waiting. Just kidding. We feel more like administrative labours than scientists.. - not kidding.

11th-12th week

We've receive the first part (one third) of our order of lab equipments. We are seeing the light in the end of the tunnel, yay! Now, we could calmly put everything into designing our oligos and synthetic genes. We could start to planning the oligos not until we were certain, that we will get the second tranche of government funds.

Semptember.

11 week

This week we've isolated the chromosome E. coli strain DH5alpha. From bacterial chromosome we want to pull out: 1. arabinose promoter 2. rhamnose promoter 3. melibiose promoter 4. xylose promoter Continously we are designing of our constructs. (redirection)

12 week

We received the first part of our oligos. We were ably to make the PCR of our promoters, backbones with proper resistances. Okey, a lot of PCR reaction with various primers. We wanted to be sure that reaction will go well, because A lot of agarose gel, as well. And photos, a lot of agarose gel photos. We are making the PCR form the AddGene plasmid, which we've ordered. On this plasmids we have: 1. phiC31 integrase 2. TP901 integrase 3. Bxb1 integrase 4. arabinose promoter

13 week

We've received the second part of oligos and synthetic genes. We are able to make the PCR reaction of fluorescent proteins: BFP, GFP and mRFP of our design, optimize for E. coli bacterium with three different tags, which should speed up the degradation of the proteins.

14 week

Let's start the Gibson assembly! Protocol you can find here. In the beginning we use the Gibson assembly kit and the Gibson assembly master mix. We've achieved nothing.