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Team:Bordeaux/NotebookAugust19
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(Created page with "{{:Team:Bordeaux/Tete}} <html> <h1>19<sup>th</sup> August </h1> <div class="type-exp"> Molecular biology</div> <br> <ul class="day-detail"> <li> Verification of our plasmid Mi...") |
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Digestion of 1μg of insert<br> | Digestion of 1μg of insert<br> | ||
| - | <b>Ex :<b> ELP = 460ng/μl<br> | + | <b>Ex :</b> ELP = 460ng/μl<br> |
<div style="margin-left:35px;">So 1μg (ELP) = 2,17μl <br>+ 7,83μl H2O<br></div> | <div style="margin-left:35px;">So 1μg (ELP) = 2,17μl <br>+ 7,83μl H2O<br></div> | ||
| - | + | <table> | |
| - | DNA | + | <tr><td>DNA </td><td> 10μl</td></tr> |
| - | Xba1 | + | <tr><td>Xba1 </td><td> 0,5μl</td></tr> |
| - | Spe1 | + | <tr><td>Spe1 </td><td> 0,5μl</td></tr> |
| - | 10X Buffer 4 5μl< | + | <tr><td>10X Buffer 4</td><td> 5μl</td></tr> |
| - | H+O | + | <tr><td>H+O </td><td> qsp 50μl</td></tr></table> |
<br> | <br> | ||
Migration on agarose 1,5% gel (100V, 30min)<br> | Migration on agarose 1,5% gel (100V, 30min)<br> | ||
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Problem with our 1x500ml Erlenmeyer bacteria grow slower than in 2x250ml erlen. | Problem with our 1x500ml Erlenmeyer bacteria grow slower than in 2x250ml erlen. | ||
</li><br> | </li><br> | ||
| - | |||
| - | |||
<li> | <li> | ||
Preculture of ELP-60 in 2x250ml of TB+amp | Preculture of ELP-60 in 2x250ml of TB+amp | ||
Latest revision as of 01:52, 18 October 2014
19th August
Molecular biology
- Verification of our plasmid Miniprep we haven’t any problems with the plasmid
-
Digestion of 1μg of insert
Ex : ELP = 460ng/μl
So 1μg (ELP) = 2,17μl
+ 7,83μl H2ODNA 10μl Xba1 0,5μl Spe1 0,5μl 10X Buffer 4 5μl H+O qsp 50μl
Migration on agarose 1,5% gel (100V, 30min)
-
Test of pSB1C3 miniprep
- Digestion : we want 1μg of plamisd
- Ligation : [insert] = 6 [plasmid] => molar concentration
- PCR: linearization of ELP-40
Molecular biology
- Problem with our 1x500ml Erlenmeyer bacteria grow slower than in 2x250ml erlen.
- Preculture of ELP-60 in 2x250ml of TB+amp
- Preculture of BL21 in 5ml of LB overnight
