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Team:Valencia/Notebook/July
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Contents |
July 8th
WETLab meeting
We start to put together all the thing, to begin working in the Lab!! We also made plans for the future.
We prepared the culture medium for growing our yeasts.
Social Event
We got together in Serranos Towers, to chill out a litle bit after the "hard work" in the lab.
July 9th
- We filled up the petri dishes with the yeast liquid culture medium.
- Then we planted the yeasts onto the petri dishes.
- We prepared an overnight culture at 37ºC into a stove with the bacteria having the plasmids pG1 and pLZ in order to purificate them next Monday.
July 10th
We transferred the bacterial culture from the stove (37ºC) to the fridge (4ºC).
July 11th
Spain won the World cup!!!!
July 12th
- We made a miniprep to purify the plasmids pG1 and pLZ.
But before you start any procedure, you have to read the protocol!!!!
- Then we carried out a digestion and the following electrophoresis to verify whether we had the constructions in the plasmids and to evaluate their concentration.
First good result!! Here a picture of the electrophoresis. From left to right: Marker, pLZ and pG1.
- We stored the non-digested plasmids in the refrigerator (we want to use it later on)
- We made (Jose made) a liquid culture of E. coli with the LEA gene, and puted it into the stove at 37ºC.
- We recieved a filter paper soaked with a plasmid from Kausik Li containing an Aplysia prion fused with the GR domain. Then we put the paper containing the plasmid in water in order to transform and clone it in E. coli later on.
July 13th
- We carried out a miniprep to purify the pM2 plasmid containing the LEA gene.
- Then we carried out a digestion and the following electrophoresis to verify whether we had the constructions in the plasmids and to evaluate their concentration.
- We performed a transformation of plasmid DNA containing a prionic gene into E.coli following a heat shock protocol.
July 14th
- We prepared 600ml of yeast minimum culture medium (SD) for our future yeast cultures.
SD Composition:
- YNB w/o a.a. .............. 6.7 g (1.7 + 5)<math>(NH_4)_2 SO_4</math>
- Glucosa ................... 20 g
- Agar ...................... 20 g
- H_2O ...................... c.s.p 1 L
-We also elaborated five amino acid and nitrogen bases solutions (Leu, His, Trp, Adenine, Uracil) for which our competent yeast strains are metabolic mutants. Solutions Composition: a.a. / b.n. ............... 100 mg H_2O d.d. ................ 50 ml
We will use them in order to differentiate between plasmid transformed and non transformed yeast colonies (Only those yeast cells carrying the plasmid with the metabolic genes that our auxotrophic mutants lack will grow).
- Finally, we re-cultured the two plasmid transformed E.coli strains (with the Aplysia and yeast prion genes) we worked the day before yesterday with.
