Team:Warsaw/Calendar-Main/20 August 2009
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(Difference between revisions)
(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3>Amplyfing of Pho sequence</h3> <h4>Justyna</h4> <p>Methods</p> <br/> <ul> <li>PCR reaction mix:</li> <br/> per 50μl: <pre> 5.0 ...) |
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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> | ||
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- | <ul> pSB plasmid was previously prepared as described <a href="https://2009.igem.org/Team:Warsaw/Calendar-Main/14_August_2009">here</a> | + | <ul><li>pSB 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 19:30, 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 pSB 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
- pSB plasmid was previously prepared as described here
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