Team:Uppsala-SwedenWeek16
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== Week-16 == | == Week-16 == | ||
+ | |||
+ | |||
+ | Start to apply high fidelity PCR due the construct are getting larger and the regular Taq polymerase may not perform as well as the Phusion Polymerase at this length. Based on the company´s manual, we calculated the | ||
+ | |||
+ | <TABLE WIDTH=405 BORDER=1 BORDERCOLOR="#000000" CELLPADDING=7 CELLSPACING=0> | ||
+ | <COL WIDTH=106> | ||
+ | <COL WIDTH=81> | ||
+ | <COL WIDTH=80> | ||
+ | <COL WIDTH=80> | ||
+ | <TR VALIGN=TOP> | ||
+ | <TD WIDTH=106> | ||
+ | <P LANG="sv-SE" CLASS="western"><BR> | ||
+ | </P> | ||
+ | </TD> | ||
+ | <TD WIDTH=81> | ||
+ | <P LANG="sv-SE" CLASS="western">Temperatur</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">Time</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">Cycle</P> | ||
+ | </TD> | ||
+ | </TR> | ||
+ | <TR VALIGN=TOP> | ||
+ | <TD WIDTH=106> | ||
+ | <P CLASS="western"><SPAN LANG="sv-SE">Initial</SPAN><SPAN LANG="sv-SE"> | ||
+ | denaturation</SPAN></P> | ||
+ | </TD> | ||
+ | <TD WIDTH=81> | ||
+ | <P LANG="sv-SE" CLASS="western">98</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">30 s | ||
+ | </P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">1</P> | ||
+ | </TD> | ||
+ | </TR> | ||
+ | <TR VALIGN=TOP> | ||
+ | <TD WIDTH=106> | ||
+ | <P LANG="sv-SE" CLASS="western">Denaturation</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=81> | ||
+ | <P LANG="sv-SE" CLASS="western">98</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">10</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">30</P> | ||
+ | </TD> | ||
+ | </TR> | ||
+ | <TR VALIGN=TOP> | ||
+ | <TD WIDTH=106> | ||
+ | <P CLASS="western"><SPAN LANG="sv-SE">Anne</SPAN><SPAN LANG="sv-SE">aling</SPAN></P> | ||
+ | </TD> | ||
+ | <TD WIDTH=81> | ||
+ | <P LANG="sv-SE" CLASS="western">55.3</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">10s</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">30</P> | ||
+ | </TD> | ||
+ | </TR> | ||
+ | <TR VALIGN=TOP> | ||
+ | <TD WIDTH=106> | ||
+ | <P LANG="sv-SE" CLASS="western">Extension</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=81> | ||
+ | <P LANG="sv-SE" CLASS="western">72</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">1 min 45 s</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">30</P> | ||
+ | </TD> | ||
+ | </TR> | ||
+ | <TR VALIGN=TOP> | ||
+ | <TD WIDTH=106> | ||
+ | <P CLASS="western"><SPAN LANG="sv-SE">Final ex</SPAN><SPAN LANG="sv-SE">tension</SPAN></P> | ||
+ | </TD> | ||
+ | <TD WIDTH=81> | ||
+ | <P LANG="sv-SE" CLASS="western">72</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">5 min</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">1</P> | ||
+ | </TD> | ||
+ | </TR> | ||
+ | <TR VALIGN=TOP> | ||
+ | <TD WIDTH=106> | ||
+ | <P LANG="sv-SE" CLASS="western"><BR> | ||
+ | </P> | ||
+ | </TD> | ||
+ | <TD WIDTH=81> | ||
+ | <P LANG="sv-SE" CLASS="western">4</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western">forever</P> | ||
+ | </TD> | ||
+ | <TD WIDTH=80> | ||
+ | <P LANG="sv-SE" CLASS="western"><BR> | ||
+ | </P> | ||
+ | </TD> | ||
+ | </TR> | ||
+ | </TABLE> |
Revision as of 19:39, 26 October 2010
Week-16
Start to apply high fidelity PCR due the construct are getting larger and the regular Taq polymerase may not perform as well as the Phusion Polymerase at this length. Based on the company´s manual, we calculated the
|
Temperatur |
Time |
Cycle |
Initial denaturation |
98 |
30 s |
1 |
Denaturation |
98 |
10 |
30 |
Annealing |
55.3 |
10s |
30 |
Extension |
72 |
1 min 45 s |
30 |
Final extension |
72 |
5 min |
1 |
|
4 |
forever |
|