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Team:Warsaw/Calendar-Main/14 August 2009
From 2009.igem.org
(Difference between revisions)
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<p>Task 1:</p> | <p>Task 1:</p> | ||
<ul> | <ul> | ||
| - | <li>Gel-out Bax PCR product</li> | + | <li>Gel-out Bax PCR product</li></ul> |
<p>Methods:</p> | <p>Methods:</p> | ||
| - | <li>Bax PCR products were isolated from agarose gel using A&A Gel-Out kit.</li></ul> | + | <ul><li>Bax PCR products were isolated from agarose gel using A&A Gel-Out kit.</li></ul> |
<br/> | <br/> | ||
<p>Task 2:</p> | <p>Task 2:</p> | ||
| Line 21: | Line 21: | ||
<p>Methods:</p><ul> | <p>Methods:</p><ul> | ||
<li>Digestion of isolated plasmids by XbaI and SpeI</li> | <li>Digestion of isolated plasmids by XbaI and SpeI</li> | ||
| - | + | <li>Reaction mixture composition: 10.0 μl purified plasmid DNA product, 0.5 μl XbaI (Fermentas),0.5 μl SpeI (Fermentas), 2 μl Buffer Orange (Fermentas), 7.0 μl MQ water</li></ul> | |
<li>The digestion was performed overnight and it was subsequently inactivated via heating in 80°C for 20 minutes.</li> | <li>The digestion was performed overnight and it was subsequently inactivated via heating in 80°C for 20 minutes.</li> | ||
<li>In the next step reaction mixtures were loaded into the agarose gel to analize restriction pattern of the plasmids and isolated using A&A Gel-Out kit</li></ul> | <li>In the next step reaction mixtures were loaded into the agarose gel to analize restriction pattern of the plasmids and isolated using A&A Gel-Out kit</li></ul> | ||
<p>Task 3:</p> | <p>Task 3:</p> | ||
<ul> | <ul> | ||
| - | <li>Ligation of Bax and pBS</li> | + | <li>Ligation of Bax and pBS</li></ul> |
| - | <p>Methods:</p> | + | <p>Methods:</p><ul> |
<li>The full description of ligation is here <a href=...></a></li></ul> | <li>The full description of ligation is here <a href=...></a></li></ul> | ||
<br/> | <br/> | ||
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<p>Task 4:</p> | <p>Task 4:</p> | ||
<ul> | <ul> | ||
| - | <li>Insertion of Bax-pBS plasmid into bacteria</li> | + | <li>Insertion of Bax-pBS plasmid into bacteria</li></ul> |
| - | <p>Methods:</p> | + | <p>Methods:</p><ul> |
<li>Previously prepared chemicompetent bacteria were used.</li> | <li>Previously prepared chemicompetent bacteria were used.</li> | ||
<li>The full description of the procedure is here <a href=...></li> | <li>The full description of the procedure is here <a href=...></li> | ||
Revision as of 11:44, 14 August 2009
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Cloning Bax into pBS plasmid
Justyna
Task 1:
- Gel-out Bax PCR product
Methods:
- Bax PCR products were isolated from agarose gel using A&A Gel-Out kit.
Task 2:
- Digestion and gel-out of pBS plasmid
Methods:
- Digestion of isolated plasmids by XbaI and SpeI
- Reaction mixture composition: 10.0 μl purified plasmid DNA product, 0.5 μl XbaI (Fermentas),0.5 μl SpeI (Fermentas), 2 μl Buffer Orange (Fermentas), 7.0 μl MQ water
Task 3:
- Ligation of Bax and pBS
Methods:
Task 4:
- Insertion of Bax-pBS plasmid into bacteria
Methods:
- Previously prepared chemicompetent bacteria were used.
- The full description of the procedure is here
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