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Team:Warsaw/Calendar-Main/15 April 2009
From 2009.igem.org
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<ul> | <ul> | ||
<li>Amplification of mgtc and hly</li> | <li>Amplification of mgtc and hly</li> | ||
| + | </ul> | ||
<p>Methods:</p> | <p>Methods:</p> | ||
| - | < | + | <ul> |
| + | <li>PCR mixture's composition: 2ul buffer*, 1ul MgCl2*, 0,5ul primers, 1,5ul dNTPs (10 mM), 01,ul polymerase**, solution topped up with H2O to 20ul. | ||
<p>2 Repeats of every sample have been made.</p> | <p>2 Repeats of every sample have been made.</p> | ||
<p>* bufor i Mg made by Grubs</p> | <p>* bufor i Mg made by Grubs</p> | ||
| - | <p>** polymerase pfu turbo from first dilution by Ziemniak</p> | + | <p>** polymerase pfu turbo from first dilution by Ziemniak</p></li> |
| + | </ul> | ||
| - | < | + | <li>PCR program</li> |
<p>- mgtc 90s 95C, 2x(30s 95C, 35s 48C, 60s 72C), 28x(30s 95C, 35s 58C, 60s 72C), 600s 72C, ~ 4C</p> | <p>- mgtc 90s 95C, 2x(30s 95C, 35s 48C, 60s 72C), 28x(30s 95C, 35s 58C, 60s 72C), 600s 72C, ~ 4C</p> | ||
<p>- hly 90s 95C, 2x(30s 95C, 35s 42C, 150s 72C), 28x(30s 95C, 35s 47C, 150s 72C), 600s 72C, ~ 10C</p> | <p>- hly 90s 95C, 2x(30s 95C, 35s 42C, 150s 72C), 28x(30s 95C, 35s 47C, 150s 72C), 600s 72C, ~ 10C</p> | ||
Revision as of 19:26, 9 June 2009
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PCR mgtc, hly
Kamil ?
Tasks:
- Amplification of mgtc and hly
Methods:
- PCR mixture's composition: 2ul buffer*, 1ul MgCl2*, 0,5ul primers, 1,5ul dNTPs (10 mM), 01,ul polymerase**, solution topped up with H2O to 20ul.
2 Repeats of every sample have been made.
* bufor i Mg made by Grubs
** polymerase pfu turbo from first dilution by Ziemniak
- mgtc 90s 95C, 2x(30s 95C, 35s 48C, 60s 72C), 28x(30s 95C, 35s 58C, 60s 72C), 600s 72C, ~ 4C
- hly 90s 95C, 2x(30s 95C, 35s 42C, 150s 72C), 28x(30s 95C, 35s 47C, 150s 72C), 600s 72C, ~ 10C
Electrophoretic separation on 1% agarose gel
Results:
Gel (from left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- mgtc 1 repeat
- mgtc 2 repeat
- mgtc sample -
- hly 1 repeat
- hly 2 repeat
- hly sample -
Notes:
Inappropriate template DNA has been used for mgtc (ganomic DNA from Yersinia instead of genomic DNA from Listeria).
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