Team:WashU/Notebook/Transformation

From 2011.igem.org

(Difference between revisions)

Latest revision as of 03:47, 28 September 2011

Transformation Group

The goal of the transformation team was to successfully integrate the four synthesized genes into the yeast genome.

Once the genes are ligated into a construct with their respective selection markers, we will use homology regions from the yeast genome to integrate the linear piece of DNA into the yeast genome. After inserting one gene per yeast cell, we will mate MATa and MATalpha strains yeast to make two diploid strains, each containing two of our genes of interest. We will then sporulate the two strains and select for a yeast strain that has integrated all four genes into its genome.

Additionally, this team transformed plasmids into e. coli in order to amplify plasmids for use by the other teams. This procedure consisted of transforming the plasmid into the bacterial genome, culturing the transformed bacteria, and isolating the DNA plasmid by means of a mini-prep kit.

@@ Line 2: / Line 2: @@
 == Transformation Group ==
-The transformation team worked on the bacterial transformation. The team took any DNA plasmid and transforms the plasmid into the bacterial genome. We then cultured the transformed bacteria and isolate the DNA plasmid. This procedure amplifies the amount of DNA plasmid.
+[[File:Yeast Transformation.jpg|300px|thumb|right|]]
-The team also transformed DNA plasmid into yeast genome. After the microbiology group ligate the genes in the plasmid, we were responsible for inserting the genes into yeast.  We were responsible for sporulation of the yeast with different gene insertions to produce daughter cells with both genes of interest.
+The goal of the transformation team was to successfully integrate the four synthesized genes into the yeast genome.
+Once the genes are ligated into a construct with their respective selection markers, we will use homology regions from the yeast genome to integrate the linear piece of DNA into the yeast genome. After inserting one gene per yeast cell, we will mate MATa and MATalpha strains yeast to make two diploid strains, each containing two of our genes of interest. We will then sporulate the two strains and select for a yeast strain that has integrated all four genes into its genome.
-===Sept. 24===
+Additionally, this team transformed plasmids into e. coli in order to amplify plasmids for use by the other teams. This procedure consisted of transforming the plasmid into the bacterial genome, culturing the transformed bacteria, and isolating the DNA plasmid by means of a mini-prep kit.
-pSB1C3 plasmid arrived; genes ligated into plasmid by MolBio group
-Transformed E. coli
+[[File:Bacteria Plate.jpg|400px|thumb|center|]]
-LB plates: 150 mL h20 + 6.25 g LB broth + 3.75 g Bacto Agar to Erlenmeyer flask
-add water to 250 mL (final volume ended up being ~130 mL)
-Chloramphenicol stock solution: 0.07 g chloramphenicol into 2 mL, filter-sterilize; add ~130 uL stock solution into LB (1:1000 dilution)