We wanted to investigate to effects of supernatant includes LALF protein to E.coli in this experiment.

Therefore we put 8 ul LB broth into falcons.then we added them E.coli which has rfp and incubated for 12 hours in 37⁰C .We put pure E.coli into only one falcon for control.We added 1ul,10ul and 100ul supernatant with LALF and without LALF.

We diluated them 7 times 2-4-8-12hours later then we spreaded 10ul to plates which has 1/0,5 l Cloramphenicol.We did santrifuge liquid culture which incubated for 12 hours at 37⁰C at 7000 rpm for 5 minutes because of preparing supernant with LALF and without LALF.Then we passed them from the 0,2 filter.

Plates were spreaded after 2 hours later:

 

1a)We added 1ul supernatant includes LALF into left plate and 1ul supernatant without LALF into right plate

 

1b)We added 10 ul supernatant includes LALF into left plate and 10 ul supernatant without LALF into right plate

 

1c)We added 100ul supernatant includes LALF into left plate and 100ul supernatant without LALF into right plate

Plates were spreaded after 4 hours later;

 

2a)We added 1ul supernatant includes LALF into left plate and 1ul supernatant without LALF into right plate.

 

2b)We added 100ul supernatant includes LALF into left plate and 100 ul supernatant without LALF into right plate.

 

 

 

Plates were spreaded after 8 hours later;

 

3a)We added 1ul supernatant includes LALF into left plate and 1ul suoernatant without LALF into right plate.

 

3b)We added 10 ul supernatant includes LALF into left plate and 10 ul supernatant without LALF into right plate.

 

3c) We added 100ul supernatant includes LALF into left plate and 100 ul supernatant without LALF into right plate.

 

 

Plates were spreaded after 12 hours later;

 

4a) We added 1ul supernatant includes LALF into left plate and 1ul suoernatant without LALF into right plate.

 

4b)We added 10 ul supernatant includes LALF into left plate and 10 ul supernatant without LALF into right plate.

 

4c)We added 100ul supernatant includes LALF into left plate and 100 ul supernatant without LALF into right plate.

	Sporocide To remove infections B.subtilis spores from our Anti-LPS factor containing biofilm paste, fenton reagent is used before use against E.coli.
	iGEM for 7 to 77 During funny summer in our project executions, we have also met hundreds of people from different ages in order to tell our project in iGEM competition and benefits of synthetic biology.
	Game We have designed monopoly like game name "The Canvas Town" to inform people from any distinct education level about features of our project, laboratory equipments and interesting Biobrick of iGEM competition.
	Canvas Times