Team:Warsaw/Calendar-Main/27 August 2009

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<p>Results:</p>
<p>Results:</p>
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<li>The intA gene that was used propably wasn't the intA gene after all. </li>
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<li>The intA gene that was used might not be the intA gene after all. </li>
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Revision as of 19:46, 6 September 2009


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Cloning of the mgtc promoter into the pSB1A3 plasmid

Kamil


Tasks:

  • Amplification of the mgtc promoter using new primers
  • Isolation of the pSB1A3 plasmid

Methods:

  • The PCR mix was prepared as follows: 

    5μl buffer B (EURx)
    5μl 10mM dNTPs
    5μl forward starter
    5μl reverse starter
    2μl OptiTaq polymerase (EURx)
     2μl Salmonella matrix
     26 μl H2O
  • PCR programme: 

     4min 95°C 
     (30s 95°C, 35s 56°C, 40s 72°C)x28 
     10min 72°C 
     ~ 4°C
  • The PCR results were visualised with gel electrophoresis on 1% agarose gel.
  • The plasmid isolation was carried out from 3μl of culture cultivated for 6h in 37°C. The Plasmid Mini kit (A&A Biotechnology) was used.

Results:

  • Nothing

Conclusions:

  • The temperature was chosen... poorly.

Cloning of the cro-box into the pSB1A3 plasmid

Kamil


Tasks:

  • Creation of the cro-box double stranded DNA

Methods:

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