Team:TU Delft/9 September 2010 content
From 2010.igem.org
Alkane degradation
Yesterday's colony PCR on colonies of 027 didn't show any positives, but seeing as there were more colonies another cPCR was run overnight with the following results: one of the colonies (F) was positive.
Lane description
# | Description | Expected Length (bp) | Correct? |
0 | Smartladder (5μl) | ||
1-10 | 027 (#A-J) | 1644bp | #F |
11 | Smartladder (5μl) |
Cell banks were made of the positive colonies (also of yesterday), and plasmids were isolated using the Birnbiom method
We also tried to make 026 again (didn't give colonies last try):
Digestion:
DNA | Buffer | BSA | Enzyme 1 | Enzyme 2 | Extra? | |
# | amount | |||||
018A | 2μl | 2 (NEB) | ✓ | EcoRI | SpeI | AseI |
019A | 2.3μl | 2 (NEB) | ✓ | EcoRI | XbaI | Phosp. |
Ligation: 10μl 018A (cut) + 10μl 019 (cut) + ligase (NEB) + ligase buffer (NEB)
Transformation: Commercial competent TOP10 cells were transformed with 2.5μl of ligation mix