Team:Warsaw/Calendar-Main/20 August 2009
From 2009.igem.org
(Difference between revisions)
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- | <h3>Cloning Pho into | + | <h3>Cloning Pho into pSB1A3 plasmid</h3> |
<h4>Justyna</h4> | <h4>Justyna</h4> | ||
<p>Task 1:</p> | <p>Task 1:</p> | ||
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<li>The arrow points the right and best isolated product</li> | <li>The arrow points the right and best isolated product</li> | ||
</ul> | </ul> | ||
- | <ul><li> | + | <ul><li>pSB1A3 plasmid was previously prepared as described <a href="https://2009.igem.org/Team:Warsaw/Calendar-Main/14_August_2009">here</a> </li> |
</ul> | </ul> |
Revision as of 21:55, 21 August 2009
Amplyfing of Pho sequence
Justyna
Methods
- PCR reaction mix:
per 50μl:
5.0 μl - 10 x buffer (20mM MgSO4) 3.5 μl - dNTP mix (5mM) 2.5 μl - primer PhoF 2.5 μl - primer PhoR2 3.75μl - DMSO 2.0 μl - template DNA 28.25μl - mQ water 2.5 μl - Yellow PfuPlus DNA polymerase (1U/μl)
Thermal cycling conditions for PCR:
94°C, 1 min 0s 94°C, 0 min 15s 55°C, 0 min 30s 68°C / 72°C (either), 1 min 0s cycles 1-25 68°C / 72°C (either), 7 min 0s 4°C, indefinite
Results:
Cloning Pho into pSB1A3 plasmid
Justyna
Task 1:
- Gel-out Pho PCR product
Methods:
- Pho PCR products were isolated from agarose gel using A&A Gel-Out kit.
- The arrow points the right and best isolated product
- pSB1A3 plasmid was previously prepared as described here
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