Team:TU-Eindhoven/Notebook/Week3

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We <span class= "red">started over from scratch</span> using the old method to get the GECOs in the yeast. We also started using another method: cloning into yeast by homologous recombination. This new method makes use of the ability of yeast to 'glue' matching pieces of DNA together. This saves us most of the hassle of the original method of molecular cloning, where we do the cutting and pasting by hand.  
We <span class= "red">started over from scratch</span> using the old method to get the GECOs in the yeast. We also started using another method: cloning into yeast by homologous recombination. This new method makes use of the ability of yeast to 'glue' matching pieces of DNA together. This saves us most of the hassle of the original method of molecular cloning, where we do the cutting and pasting by hand.  
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On the plate with 200uL of transformants, we <span class= "red">finally got colonies</span>! But perhaps too many this time... Therefore we made new plates with 20uL and 2uL of transformants. The 20uL plates show good results, while the 2uL plates were empty. We did colony PCR and put the useful colonies on another plate. We tested 10 colonies of each color on the gel. The results: 4 red colonies, 0 green colonies and 1 blue colony.
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On the plate with 200μL of transformants, we <span class= "red">finally got colonies</span>! But perhaps too many this time... Therefore we made new plates with 20 μL and 2 μL of transformants. The 20 μL plates show good results, while the 2 μL plates were empty. We did colony PCR and put the useful colonies on another plate. We tested 10 colonies of each color on the gel. The results: 4 red colonies, 0 green colonies and 1 blue colony.

Latest revision as of 01:44, 27 September 2012