Monday |
Tuesday |
Wednesday |
Thursday |
Friday |
Saturday |
Sunday |
30/06
- PCR lasR (BBa_C0079). The purpose of this PCR was to add the RBS (BBa_K143021), through addition of the sequence in the primer foward, and also to remove the LVA tag. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this part will be called BBa_K1521001.
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01/07
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02/07
- Transformation of the Biobricks:
- Inoculum of the Biobricks sent by iGEM HQ:
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03/07
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04/07 |
05/07 |
06/07
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07/07
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08/07 |
09/07
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10/07
- Miniprep, Quantification and Restriction Analysis:
- BBa_B0015 (restriction analysis fail, repeat)
- PCR the qteE gene for amplification. We also added the RBS (BBa_K143021,) using the foward primer. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this biobrick will be called BBa_K1521000.
- Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)
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11/07
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12/07 |
13/07 |
14/07
- Assembly:AIII
- Prepare X-gal
- Prepare more LB medium (solid + liquid)
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15/07
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16/07
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17/07
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18/07
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19/07 |
20/07 |
21/07
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22/07
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23/07
- Assemblies AIII and KI:
- Prepare more competent cells
- Prepare LB medium (solid and liquid)
- Inoculum
- Prepare RBS+lasR and RBS+qteE for sequencing
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24/07
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25/07
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26/07 |
27/07 |
28/07
- Assembly AIII:
- Restriction analysis (+NdeI enzyme)
- OBS: This analysis was reapeted because we had problems confirming the assembly. The issue was that both the construction and the pSB1C3 vector had similar size.
- Assembly AII,
AV and
AVI:
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29/07
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30/07
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31/07
- Assembly AII,
AV and
AVI:
- PCR BBa_K143055 for RBS removal
- OBS: This was one of the parts sent by the Imperial College Team. Because of a comunication failure we assumed that this part was comming without the RBS, so we included a RBS in the parts we were going to use with this Lac promotor :)
- PCR of Plac BBa_K143055 was followed by a Gel Purification and Ligation in the PUC19 vector
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