Team:Technion/19 September 2012

From 2012.igem.org



Team:Technion/19_September_2012
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Ilya

- Put starters of the parts I worked on. These starters will be used for minipreping the liquid DNA for parts submission.
- Discussed with Roee the presentation of the riboswitch results.

Inbal

-Gel purification for T3 RNAP (2700bp): 317ng/ul.
-miniprep for In1: 196, 161.5 and 169.5 ng/ul.
-miniprep for In1: 227.5, 288.5, 209.5 mg/ul.
-The restiction I made today:
1.pSB1C3 with XbaI.
2.In1 with XbaI and SpeI. After running the products on agarose gel, there were 2 bands at 3kb approximately, I assumed it was the backbone and T7 polymerase, but not for sure, because the bands were too close to each other (maybe I don't have the whole desired clone, but I will check it later). After that I, digested In1 with XbaI and ran it on gel, the band was 6.8kb, but not for sure... :/
3.T3 with XbaI and SpeI.
- cip reaction for pSB1C3.
- starter for In1 (for glycerol stock).
-I ligated T3 RNAP and pSB1AK3, and then transformed it to ROP10 bacteria.
- Transformation of In1+xyIE and In1+ALP to TOP10 bacteria.

Asaf

I lygated the different inserts with the psb1c3 backbone.

Hila

- Gel run for all the PCR products. Shahar will gel – purifies it, and cut the fragments with PacI and SpeI restriction enzymes.
- HR ladder tests. We found that the ladder does not open as it should, so I started testing the appropriate conditions for this ladder.

Lior

preparing the shipping plasmids for our parts.

Noa

preparing the shipping plasmids for our parts.

Evgeni

Shahar

gel purified all PCR products, and cut the fragments with PacI and SpeI restriction enzymes

Rachel

Asaf did 12 starter, 4 for each RNAPs (4 clones of K1F, 4 clones of N4, 4 clones of T3).