| 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.
|
01/07
|
02/07
- Transformation of the Biobricks:
- BBa_143012
- BBa_K081001
- BBa_E0840
- Inoculum of the Biobricks sent by iGEM HQ:
- BBa_K316037
- BBa_K316018
- BBa_K316015
|
03/07
|
04/07 |
05/07 |
06/07
- Inoculum:
- BBa_143012
- BBa_K081001
- BBa_E0840
|
| 07/07
|
08/07 |
09/07
|
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)
|
11/07
|
12/07 |
13/07 |
14/07
- Assembly:AIII
- Prepare X-gal
- Prepare more LB medium (solid + liquid)
|
15/07
|
16/07
|
17/07
|
18/07
|
19/07 |
20/07 |
| 21/07
|
22/07
|
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
|
24/07
|
25/07
|
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 bot the construction and the pSB1C3 vector had similar size. Check out the Lab Journal report of the day for more information.
- Assembly AII,
AV and
AVI:
|
29/07
|
30/07
|
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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