Team:Warsaw/Calendar-Main/24 September 2008

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Revision as of 22:48, 13 October 2008


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Results of mutagenesis: no colonies on any plate.
Mutagenesis repeated with modified conditions: fresh stock of dNTPs used and additional MgCl2 added.

PCR on pKS plasmid containing protein A with AL+link10+homo2 and AP+NotI primers (20 cycles, elongation duration 15 s, annealing temperature 72°C).
PCR on pUC19 plasmid with AlphaL+SacI and AlphaP+link10+homo2 primers (20 cycles, elongation 45 s, annealing temperature 63°C).
As a result we got two PCR products (alpha-linker and linker-A) wich will be utilized as templates in next PCR.
Gel electrophoresis of PCR products and gel-out of proper bands (alpha_linker - 572 bp and linker_A - 467 bp ).
Measurment of concentration of both isolated products

@@ Line 10: / Line 10: @@
-<h3>Preparation of alpha+A construct</h3>
+<h3>Preparation of alpha_A construct</h3>
 <h4>Antoni</h4>
 <p>
@@ Line 19: / Line 19: @@
 <li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=http://www.fermentas.com/techinfo/nucleicacids/mappuc1819.htm>pUC19</a> plasmid with
 <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaL+SacI">AlphaL+SacI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaP+link10+homo2">AlphaP+link10+homo2</a>
-  primers (20 cycles, elongation duration 45 s, annealing temperature 63&deg;C).
+  primers (20 cycles, elongation 45&nbsp;s, annealing temperature 63&deg;C).
 <br>
 As a result we got two PCR products (alpha-linker and linker-A) wich will be utilized as templates in next PCR. </li>