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Team:TUDelft/30 July 2009
From 2009.igem.org
Lab Notebook
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30 July 2009
Tim Weenink
prepared assembly mixes for !B, !C and !D constructs. Standard Ginko assembly kit was used. DNA concentrations as below (these mixtures were added to 7.5 µl of digestion mixture):
| Assembly name | position | Components | volume in µl |
| !B | Upstream | !A DNA | 33.0 |
| H20 | 9.5 | ||
| Downstream | BBa_B0015 DNA | 10.0 | |
| H20 | 32.5 | ||
| Backbone | pSB1AC3 DNA | 6.5 | |
| H20 | 36 | ||
| !C | Upstream | BBa_R0040 DNA | 10.0 |
| H20 | 32.5 | ||
| Downstream | *I6 DNA | 5.5 | |
| H20 | 37.0 | ||
| Backbone | pSB1AC3 DNA | 6.5 | |
| H20 | 36 | ||
| !D | Upstream | *I6 DNA | 5.5 |
| H20 | 37.0 | ||
| Downstream | BBa_K145201 DNA | 18.0 | |
| H20 | 24.5 | ||
| Backbone | pSB1AC3 DNA | 6.5 | |
| H20 | 36 |
Ligated and transformed assemblies according to protocol.
15:20 Done plating the assemblies.
15:40 inoculated *S and *T cultures, four colonies each
16:55 Inoculated !A cultures from glycerole stock. Two tubes.
Calin
Plated three backbones left over from the transformation the other day. Did a digest on oriT-R with E and P.
Tim Vos
Minipreped 4 x pSB1C3, 4 x pSB4A5, 2 x rbs-GFP, and 4 x pSB1A3. One pSB4A5 tube is incorrect, it shows RFP expression. Also loaded the gel.
Orr
Used the competent cell protocol to make cells with TSS competent. Started making a Java program that will allow lock sequence design based on the chosen RBS, and will design the equivalent key based on the lock sequence. The program will incorporate a database with all the working RBS segments in the iGEM registry, so that the user can choose which RBS he wants to use.
