From 2012.igem.org
Team:Technion/24_September_2012
Ilya
- wiki, wiki, wiki...
Inbal
Asaf
-I ran the colony PCR products on gel in order to establish if the insert got in to to plasmid.
I only got posotive results for the RS+N4 RNAP and RS+T7 RNAP.
-I minipreped them and send them to the regestry as BioBricks.
Hila
I performed yesterday's experiment again, this time trying to assemble three parts only: F4+F5+F6.
I created five reactions:
- 2/3 of the DNA amount as yesterday (~100ng of each fragment) for 1 hour.
- Same DNA amount as yesterday (~150ng of each fragment) for 2 hour.
- Same DNA amount as yesterday (~150ng of each fragment) for 3 hour.
- Same DNA amount as yesterday (~150ng of each fragment) for 1 hour, with a NEB Gibson solution addition.
- Same DNA amount as yesterday (~150ng of each fragment) for 2 hour, with a NEB Gibson solution addition.
We saw that the reaction time does not influence the reaction efficiency. I still couldn't assemble the three parts together.
In addition, Shahar assembled F1+F2 and F3+F4 fragments, in order to gel purified the products and try to reassemble them together into a 4 fragments sequence. His results weren't as good as shown in the previous experiment, and got us to think that they may have a problem with the T5 exonuclease (that happened due to freezing and de-freezing it).
We will create the T5 stocks again, and this time we will divide it into smaller stock so every stock will be enough for one reaction only.
We will try again this reaction with the new T5-exo stock.
Lior
Noa
Evgeni
Shahar
I assembled F1+F2 and F3+F4 fragments, in order to gel purified the products and try to reassemble them together into a 4 fragments sequence.The results weren't as good as shown in the previous experiment, and got us to think that they may have a problem with the T5 exonuclease (that happened due to freezing and de-freezing it).
Rachel
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