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Team:Warsaw/Calendar-Main/22 April 2009
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<p>Methods:</p> | <p>Methods:</p> | ||
| - | <ul><li><p>PCR mixture's composition:</p> 2,5μl pfu buffer (Fermentas), 2, | + | <ul><li><p>PCR mixture's composition:</p> 2,5μl pfu buffer (Fermentas), 2,5μl MgSO4 (Fermentas), 1,5μl primers, 1,5μl dNTPs (10 mM), 0,5μl pfu turbo polymerase <font color="green">(od Antka, ale KNGiE też powinna działać)</font>, 1μl template DNA from Listeria, solution was topped up with H2O to 25μl. |
<p>1 repeat of every sample was made (2 different programs).</p> | <p>1 repeat of every sample was made (2 different programs).</p> | ||
<p>Additionally for each sample was made version with 10X dilution of template.</p> | <p>Additionally for each sample was made version with 10X dilution of template.</p> | ||
Revision as of 11:14, 12 July 2009
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PCR hly
Kama
Tasks:
- Amplification of hly
Methods:
PCR mixture's composition:
2,5μl pfu buffer (Fermentas), 2,5μl MgSO4 (Fermentas), 1,5μl primers, 1,5μl dNTPs (10 mM), 0,5μl pfu turbo polymerase (od Antka, ale KNGiE też powinna działać), 1μl template DNA from Listeria, solution was topped up with H2O to 25μl.1 repeat of every sample was made (2 different programs).
Additionally for each sample was made version with 10X dilution of template.
- PCR programs:
hly
4min 95°C
(30s 95°C, 35s 42°C, 1min20s 72°C)x3
(30s 95°C, 35s 47°C, 1min20s 72°C)x28
10min 72°C
~ 7°C
random program
- Electrophoretic separation on 1% agarose gel
Results:
- Gel (from left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- hly control -
- hly
- hly (10x diluted template)
- hly control -*
- hly*
- hly (10x diluted template)*
* random program
hly was succesfully amplified (termocykler z gradientem w pokoju 145)
