Team:Warsaw/Calendar-Main/15 August 2009
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3><div style="text-align: center;">Assembly of endosomal detection operon</div></h3> <h4>Marcin</h4> <br/> <p>Task 1:</p> <ul><li>Ge...) |
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<li>Gel-outs were prepared according to manufacturer's manual</li></ul> | <li>Gel-outs were prepared according to manufacturer's manual</li></ul> | ||
<p>Results:</p> | <p>Results:</p> | ||
+ | <center><img src="https://static.igem.org/mediawiki/2009/f/f7/Control_gel_out_15_08_09.png" width="60%" heigth="60%"></center><font face="Times New Roman" size="3"><p><div style="text-align: center;">Evaluation of gel-outs yield</div></p></font> | ||
+ | <p><b>Comment:</b></p> | ||
<p>Some purification were unsuccessful because time required to efficacious activation of the columns is, in fact longer than few minutes (which is not depicted in the manual). It is obligated to repeat digestion and gel-out for the samples which were not isolated</p> | <p>Some purification were unsuccessful because time required to efficacious activation of the columns is, in fact longer than few minutes (which is not depicted in the manual). It is obligated to repeat digestion and gel-out for the samples which were not isolated</p> | ||
<p>Task 2:</p> | <p>Task 2:</p> |
Latest revision as of 21:09, 13 September 2009
15 August 2009
Assembly of endosomal detection operon
Marcin
Task 1:
- Gel-out of following sequences from the pSB1A3 plasmid:
Methods:
- Gel-outs were prepared according to manufacturer's manual
Results:
Evaluation of gel-outs yield
Comment:
Some purification were unsuccessful because time required to efficacious activation of the columns is, in fact longer than few minutes (which is not depicted in the manual). It is obligated to repeat digestion and gel-out for the samples which were not isolated
Task 2:
- Ligation of following constructs:
Methods:
- Ligation mix:
1.0 μl - T4 ligase 2.5 μl - Tango buffer (Fermentas, 10x) 3.0 μl - dNTPs (EURx, 5μlM) 6.0 μl - digested vector 12.5 μl - digested insert
Task 3:
- Prepare the bacterial cultures for isolation of following plasmids:
Task 4:
- Transformation of chemocompetent E. coli strain TOP10 to reveal the quality of prepared bacteria
Plasmid to transform: pSB1A3
Methods:
- Detailed protocol of transformation is described here.
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