Team:Warsaw/Calendar-Main/20 August 2009
From 2009.igem.org
(Difference between revisions)
Line 62: | Line 62: | ||
<p>Methods:</p> | <p>Methods:</p> | ||
<ul><li>Pho PCR products were isolated from agarose gel using A&A Gel-Out kit.</li> | <ul><li>Pho PCR products were isolated from agarose gel using A&A Gel-Out kit.</li> | ||
+ | <br> | ||
<center> | <center> | ||
<img src="https://static.igem.org/mediawiki/2009/3/39/Phoafergeloutopis.jpg"></center> | <img src="https://static.igem.org/mediawiki/2009/3/39/Phoafergeloutopis.jpg"></center> | ||
+ | <br> | ||
<li>The arrow points the right and best isolated product</li> | <li>The arrow points the right and best isolated product</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
|
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
|