IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/08/22
|Week Twelve|| Main project page|
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Diluted soil samples 79-84 from 10^-1 to 10^-4 and plated 10^-2 to 10^-4 on SFM + Ny.
Set up 38 more Bioassays from spore stocks - spotted 2 x 5 ul of spore stock onto a single SFM plate. The plates were left to grow at 30 ° for 7 days.
Set up overnight ETpuz cultures for tomorrow's conjugation. Added 50 ul E. coli cells to ~ 10 ml LB (lacking NaCl for ETpuz pMS82 cells as Hygromycin is salt-intolerant). Added 10 ul Chloramphenicol and Kanamycin to both ETpuz pAU3-45 and pMS82 cultures, and 10 ul of Apr for pAU3-45 and 10 ul of Hyg for pMS82.
We discovered after analysing the sequencing data from 18/07 that sample 3a had the correct sequence for biobrick BBa_K0141000 but sample 3b did not. This explained why sample 3b2, which was a miniprep from 3b, had extra NdeI sites. To confirm our theory, samples 3a, 3b, 3c and the original BBa_J04450 were cut with restriction enzyme pvuII and analysed by gel electrophoresis fig 3.
The registration for the jamboree was completed for the team.