Team:Todai-Tokyo/Notebook/bread

From 2009.igem.org

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*TA cloning of mtlD<BR>
*TA cloning of mtlD<BR>
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=='''9/21'''==
*Miniprep of mtlD<BR>
*Miniprep of mtlD<BR>

Revision as of 03:36, 16 October 2009

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Contents

Plan

Aim:Create yeast that can be used to make sweet and low energy bread

Methods:

  1. Clone the glucoamylase gene from Saccharomycopsis fibuligera and insert it in the yeast chromosome by homologous recombination
  2. Clone mtlD (mannitol synthase) from E.coli and insert it in the yeast chromosome by homologous recombination
  3. Replace gpd1 gene by Glu1 and gpd2 gene by mtlD

~9/20

  • PCR of mtlD
  • PCR of gpd1 promoter
  • TA cloning of mtlD

9/21

  • Miniprep of mtlD

9/22

  • read the sequence of mtlD→successful
  • mtlD primers with HAtag come

9/25

  • PCR of mtlD with new primer→failed

9/26

  • PCR of mtlD with new primer→successful

9/27

  • PCR of gpd1 promoter with Pfu Ultra

October

  • PCR of Glu1
  • TA cloning of Glu1
  • PCR of gpd1 promoter with ExTaq

10/14

  • cut mtlD and plate1 7D both by EcoRI and PstI
  • colony PCR of Glu1






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