Team:Warsaw/Calendar-Main/29 June 2009
From 2009.igem.org
PCR Llo
Kuba
Tasks:
- Amplification of llo(hly) for fusion with the secretion signal peptide
Methods:
PCR mixture's composition:
2,5ul pfu buffer (Fermentas), 2,5ul MgSO4 (Fermentas), 1,5ul primers, 1,5ul dNTPs (10 mM), 0,5ul pfu turbo polymerase, 1ul template DNA from Listeria, solution was topped up with H2O to 25ul.One of the samples also contained 1 ul of DMSO
- PCR programs:
hly
4min 95°C
(30s 95°C, 40s 43°C, 1min30s 72°C)x3
(30s 95°C, 40s 45°C, 1min30s 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)
- sample with DMSO
- sample without DMSO
no products were observed
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