Team:UCL London/Week 5

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UCL IGEM 2010

RETURN TO IGEM 2010

Preparation week - Week 5

26th July - 1st August

UCL-Plate1.JPG

We started transformation of the DNA we need from the iGEM registry plates today. WE had 11 parts to transform, so Xiang and the team had quit a tough few days ahead of them. We proceeded by carrying out a transformation Transformation from the iGEM plates of the parts required, 11 to be precise, with the results expected the following morning

 
UCL-Plate2.JPG

The results of the transformation on the previous day were confirmedd today...the e.coli cells clearly are not fond of us. Plate 9 (pSB1T3) had no growth on agar plate, and also ontrol plate for chloramphenicol had growth.

• Pick 2 different colonies from each plate; inoculate in 2ml LB+antibiotics overnight (O/N). • Make new 1000X chloramphenicol stock. • Transformation of part 15,16,17.


 
UCL-Gel.jpg


• Miniprep & nanodrop (to measure DNA concentration) • Run a diagnostic gel on obtained DNA • Repick 2 (C-G), 5(C-G), 6(C-G), 8(C-G), 11(C-G), together with plate 15(A&B) & 17(A&B).


Gel Result: Load from left to right as follows; HyperLadder I was used as marker. L 1A 1B 2B 3A 3B 4B L 6A 6B 7A 7B 8B 10A 10B


In summary, 2B & 4B had no DNA. 10A contained wrong plasmid or genomic DNA. Results on next day: 2-6 had no growth in 2ml LB.

Clearly, it looked like it'll take some time for us to get fully acquanted with the e.coli cells, Results 2-6 had no growth in 2ml LB. Maybe if we called ouirselves "don colioness" they might start showing some respect, maybe,....we'll see.

 
UCL-meetingportico.JPG

This Thursday meeting started as a one time event..and became like a bad habit..but in a good way. The portico seams to provide a very inspiring atmosphere for the iGEM UCL members to discuss their weekly progress and the tasks assigned to each one. For once more, a lot of ideas regarding the project were circulated.

Miniprep of 8DEF, 15AB, 17AB, & 11D. Transformation of part 13, 14, 18-23. Pick colonies from plate 2, 5, 6; grow in LB and TB.


 
UCL-Xianglab.jpg

Ahaa, the don coliones it is....the results of yesterdays experiments were out;

2C (TB), 5E-5G (TB), 6E-6G(TB), 6D (LB) --- growth. Plates:

13 Plate √ 14 Plate √ 18 Plate √ 19 X 20 Plate √? (~ 5 dark colonies) 21 X 22 X 23 X

we have the following parts (confirmed by gel electrophoresis):

Inducible promoter lacI

Possible hypoxia promoter from UCL iGEM 2009 mNarK. Positive feedback loop promoter, LasR regulated. Terminator. Kanamycin backbone for assembly. Chloramphenicol backbone for assembly. Amp&Kana backbone as back-up plan for assembly. Amp&Chlor backbone as back-up plan for assembly.


The missing important parts we need for ligation

RBS + coding region of LasR + terminator Red dye pathway (or any other colour) Native hypoxia promoter NarK.

 


Week 6

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