IGEM:Cambridge/2008/Notebook/Bacillus/2008/08/08: Difference between revisions

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|  No DNA || Spc50 || 100μL || about 12, maybe more : problem!!!!!
|  No DNA || Spc50 || 100μL || about 12, maybe more : problem!!!!!
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==Result for test of ECE112 transformation==
On Cm5 plates, we have some colonies for each strains, on Cm5 + Spc 100 plates, no colonies on each plates. This result seems good, however, we forgot to make a control. Since, we had some colonies on control plates, it is possible that our bacillus are resistant to Cm5, even if they are not transformed.
We addded some control colonies.
==Control with erythromycin==
- Prepare erythromycin, stock 5mg/mL : 0.05g in 10mL of SDW
- Prepare plates : add 20μL of Ery (5mg/mL) into 200mL of agar (final concentration : 0.5μg/mL)
- Do several spots on each plate
{|class="wikitable" style="text-align:center" border="1"
|-
!  Plate !! Colonies added from
|-
|1 || ECE166 + 100μL of cells (2 different plates) 
|-
|2|| ECE153 + 100μL of cells (2 different plates)
|-
|3 || ECE166 (from glycerol stock)
|-
|4 || Control (from a plate of IA771 without antibiotic)
|-
|5 || IA771 + ECE112 (05/08)
|-
|6 || IA771 + ECE166 (05/08)
|}
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==Amylase production screening==
* Prepare agar plate with starch
- Starch : 1g of starch in 10mL of SDW
- Add 2mL of Starch solution into 200mL of agar, mix
-Inoculate on different plates : 1A1 +ECE112 (plate from 05/08), IA751 + ECE112 (plate from 05/08) and IA771 + ECE153
==New stocks==
- Take 1mL from LB stock oh ECE171 (from 07/08) and put it into 99mL of LB
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__NOTOC__
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Revision as of 08:19, 5 September 2008

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Result from yesterday transformation

Vector Antibiotic Quantity of cells added number of colonies
ECE166 Cm5 200μL 0 : problem!!!
ECE166 Cm5 100μL 2
ECE166 Cm5 50μL 1
ECE166 Cm10 100μL 0 : no resistance to Cm10!
ECE166 (spin) Cm5 100μL a lot, confluent
ECE166 (spin) Cm10 100μL 0
ECE166 (glycerol + spin) Cm5 100μL about 150, very small
ECE166 (spin) Cm10 100μL 0
ECE153 Spc50 200μL 21 + a lot of confluent colonies
ECE153 Spc50 100μL 7 + confluent (a few)
ECE153 Spc50 50μL 4
ECE153 (spin) Spc50 100μL 25 + about 200 (small and almost confluent)
No DNA Cm5 100μL 0
No DNA Cm10 100μL 0
No DNA Spc50 100μL about 12, maybe more : problem!!!!!

Result for test of ECE112 transformation

On Cm5 plates, we have some colonies for each strains, on Cm5 + Spc 100 plates, no colonies on each plates. This result seems good, however, we forgot to make a control. Since, we had some colonies on control plates, it is possible that our bacillus are resistant to Cm5, even if they are not transformed. We addded some control colonies.

Control with erythromycin

- Prepare erythromycin, stock 5mg/mL : 0.05g in 10mL of SDW

- Prepare plates : add 20μL of Ery (5mg/mL) into 200mL of agar (final concentration : 0.5μg/mL)

- Do several spots on each plate

Plate Colonies added from
1 ECE166 + 100μL of cells (2 different plates)
2 ECE153 + 100μL of cells (2 different plates)
3 ECE166 (from glycerol stock)
4 Control (from a plate of IA771 without antibiotic)
5 IA771 + ECE112 (05/08)
6 IA771 + ECE166 (05/08)

Amylase production screening

  • Prepare agar plate with starch

- Starch : 1g of starch in 10mL of SDW

- Add 2mL of Starch solution into 200mL of agar, mix

-Inoculate on different plates : 1A1 +ECE112 (plate from 05/08), IA751 + ECE112 (plate from 05/08) and IA771 + ECE153

New stocks

- Take 1mL from LB stock oh ECE171 (from 07/08) and put it into 99mL of LB



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