IGEM:Cambridge/2008/Notebook/Bacillus/2008/08/12: Difference between revisions
(Autocreate 2008/08/12 Entry for IGEM:Cambridge/2008/Notebook/Bacillus) |
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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. | |||
Revision as of 08:30, 5 September 2008
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Results from yesterday
- 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
- IA771 : no growth on Spc50 plate, so no natural resistance - transformed cells grew
- 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 amylaseWe 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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