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Team:Warsaw/Calendar-Main/30 April 2009
From 2009.igem.org
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Cloning of hly gene into pKSII+ vector
Kamil
Tasks:
- Verify the correctness of the pKS/hly constructs
Methods:
- The BanI enzyme was selected for the test because the hly insert bears an additional digest site.
Digest mixture's composition:
2ul Orange buffer (Fermentas) 1ul BanI enzyme 1ul plasmid solution
The solution was topped up with H2O to 20ul.- The digest was kept at 37 °C overnight (~18h) and then inactivated at 65 °C for 10 min.
- The electrophoretic separation was carried out on 1% agarose gel
Results:
- Gel (from left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas) (3ul)
- sample no. 15 (5ul)
- sample no. 16 (5ul)
- sample no. 17 (5ul)
- sample no. 18 (5ul)
- pKS plasmid (control) (5ul)
Notes:
- At least 5ul of plasmid solution should have been used
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