Team:Mexico-UNAM-CINVESTAV/Notebook/Week Five
From 2010.igem.org
Week #5
04th October - 08 October 2010
Monday
- Let's picked up colonys our transformant cells and prepare all to do miniprep.
For this we used 30ml de LB liquid agar medium per falcon tube at 80μ/ml
Cloramphenicol concentration we left overnight culture to 37º
Tuesday
With a overnight inoculum we did a miniprep.
The miniprep was done with a Quiagen Kit.
Then we going to do a duble digest with EcoRI and PstI to check out our ligations.
DNA | 5μl |
Buffer NB2 | 1μl |
EcoRI | 0.5μl |
PstI | 0.5μl |
H2O | 2.7μl |
BSA | 0.3μl |
Total | 10μl |
Wensday
We run a gel to confirm our ligations as follow
- PCR 1 Lig S with Psb1C3 Confirmed
- PCR 2 Lig S with Psb1C3 Unconfirmed
- PCR 3 Lig S with Psb1C3 Confirmed
- PCR 4 Lig C with Psb1C3 Unconfirmed
![Right](/wiki/images/0/03/Con1.gif)
We prepare all for repeat ligation PCR2 with Psb1C3 and PCR4 with Psb1C3