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Team:Warsaw/Calendar-Main/2 August 2009
From 2009.igem.org
(Difference between revisions)
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<p>Task 1:</p> | <p>Task 1:</p> | ||
<ul> | <ul> | ||
| - | <li>Prepare PCR reaction to | + | <li>Prepare PCR reaction to amplifie p53 coding sequence.</li> |
</ul> | </ul> | ||
<p>Methods:</p> | <p>Methods:</p> | ||
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<li>DNA template dilution:</li> | <li>DNA template dilution:</li> | ||
</ul> | </ul> | ||
| - | <p>1 & | + | <p>1 μl of plasmid solution (isolated in 11.07.09) was taken and diluted with 99 mi&;l of MQ water</p> |
<ul> | <ul> | ||
<li>PCR mixture composition:</li></ul> | <li>PCR mixture composition:</li></ul> | ||
Revision as of 19:58, 13 August 2009
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Cloning of p53 coding sequence
Marcin
Task 1:
- Prepare PCR reaction to amplifie p53 coding sequence.
Methods:
- DNA template dilution:
1 μl of plasmid solution (isolated in 11.07.09) was taken and diluted with 99 mi&;l of MQ water
- PCR mixture composition:
- proper mixture 1: 0.5 μl primer 1 (50 nM; Oligo.pl), 0.5 μl primer 2 (50 nM; Oligo.pl), 2.5 μl dNTPs (20 μM ;Fermentas), 2 μl Pfu polymerase (EurX), 5 μl Pfu Buffer (EurX), 1 μl DNA template, 39 μl MQ water
- Negative control: the same as proper mixture 1, the only distinction is lack of the DNA template.
- Program:
p53 (detailed destription is here)
Task 2:
- Prepare the bacterial cultures for isolation of plasmids containing ligated biobricks
Preparation of bacterial cultures
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