IGEM:Cambridge/2008/Notebook/Bacillus/2008/08/12

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Results from yesterday

 * Ery plates

- IA751 (control) : most of colonies did not grow, which is good, but 2 or 3 grew...

- IA7771 + ECE166 : all colonies grew : ok 9non integration vector)

- IA771 + ECE153 : growth of all colonies... problem!!! if IA771 is transformed with this integration vector, it should lose its EryR

- IA771 + ECE112 : growth of all colonies... problem!!! if IA771 is transformed with this integration vector, it should lose its EryR


 * Spc resistance

- IA771 : no growth on Spc50 plate, so no natural resistance

- transformed cells grew


 * Amylase plates

- Add iodine solution for 1min

- Problem : no aparent blue, no diffusion of iodine into LA agar + a lot of contamination

New plates and LB stocks
- In order to try to make plasmid miniprep at the end of the day, grow IA771 transformed with ECE166 in 10mL LB + Cm5

7hours is not enough to reach exponential phase!!! We will try again tomorrow by incubating longer

- Grow IA771 and IA751 ( first plate from sterile disks of strain) imto 10mL of LB

- Grow ECE166, 171, 153 in E.coli with approriate antibiotics 9to transform tomorrow)

- Grow IA771 transformed with ECE153 into LB+Spc50 (to check fluorescence with xylose induction)

New test for amylase
We tried 2 different methods.

- Dilute 1g of starch into 100mL of agar and try to dilute it and boil it to sterilize. The problem is that it is very difficult to dilute...

- The second method seem to be better. We diluted 1g of starch into 100mL of Soft Agar (it dilutes very well). Then plate blank agar plate (LA) and then add a thin layer of SA ith 1% of starch. Poke the plates.

- We plated IAI+ECE112, IA751+ECE112 and also IA751 and 1A1 for control.


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