Team:Warsaw/Calendar-Main/30 April 2009

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)</li> </ul> <ol> GeneRuler DNA Ladder Mix #SM0333 (Fermentas) (3ul)</li> sample no. 15 (5ul)</li> sample no. 16 (5ul)</li> sample no. 17 (5ul)</li> sample no. 18 (5ul)</li> pKS plasmid (control) (5ul)</li>

</ol>

Notes:  At least 5ul of plasmid solution should have been used</li> </ul>