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Team:The University of Alberta/17 July 2008
From 2008.igem.org
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*More Colony PCR on the synthed biobricks. The final one has arrived! | *More Colony PCR on the synthed biobricks. The final one has arrived! | ||
*Did sequencing for BisDA to cofirm whether it is the part, because the colony PCR was not as expected. '''EDIT''': The colony PCR was fine; parts were a bit larger than we had thought because we did not account for the MCS being PCR'd along with the parts because of the primers we used. | *Did sequencing for BisDA to cofirm whether it is the part, because the colony PCR was not as expected. '''EDIT''': The colony PCR was fine; parts were a bit larger than we had thought because we did not account for the MCS being PCR'd along with the parts because of the primers we used. | ||
| - | *Digested Tryp with Eco/Spe (sequential digest; first with Spe and then with Eco) and J6 with Eco/Xba. This will allow us to ligate the two together. | + | *<strike>Digested Tryp with Eco/Spe (sequential digest; first with Spe and then with Eco) and J6 with Eco/Xba. This will allow us to ligate the two together.</strike> It's easier to digest both with Xba and Pst. In the case of adding inserts to vectors THAT ALREADY CONTAIN PARTS, it would be better to digest the inserts with Xba and Pst, and vectors with Spe and Pst to add the new insert behind the one already in the vector. |
Revision as of 17:05, 18 July 2008
Today
- Did Minipreps of I0500 in the new vector: Now we have I0500 in a high copy plasmid! No more adding IPTG to switch it from low-copy to high-copy mode!
- More Colony PCR on the synthed biobricks. The final one has arrived!
- Did sequencing for BisDA to cofirm whether it is the part, because the colony PCR was not as expected. EDIT: The colony PCR was fine; parts were a bit larger than we had thought because we did not account for the MCS being PCR'd along with the parts because of the primers we used.
Digested Tryp with Eco/Spe (sequential digest; first with Spe and then with Eco) and J6 with Eco/Xba. This will allow us to ligate the two together.It's easier to digest both with Xba and Pst. In the case of adding inserts to vectors THAT ALREADY CONTAIN PARTS, it would be better to digest the inserts with Xba and Pst, and vectors with Spe and Pst to add the new insert behind the one already in the vector.
