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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> | ||
| - | DNA 10μl<br> | + | DNA <div style="margin-left:35px;">10μl<br></div> |
| - | Xba1 0,5μl<br> | + | Xba1 <div style="margin-left:35px;">0,5μl</div><br> |
| - | Spe1 0,5μl<br> | + | Spe1 <div style="margin-left:35px;">0,5μl</div><br> |
| - | 10X Buffer 4 5μl<br> | + | 10X Buffer 4 <div style="margin-left:35px;">5μl</div><br> |
| - | H+O qsp 50μl<br> | + | H+O <div style="margin-left:35px;">qsp 50μl</div><br> |
<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> | ||
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| - | |||
<li> | <li> | ||
Preculture of ELP-60 in 2x250ml of TB+amp | Preculture of ELP-60 in 2x250ml of TB+amp | ||
Revision as of 01:50, 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) = 2,17μlDNA
+ 7,83μl H2O10μlXba10,5μl
Spe10,5μl
10X Buffer 45μl
H+Oqsp 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
