IGEM:Cambridge/2008/Notebook/Bacillus/2008/09/03

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Result from bacillus transformation with fresh competent cells (02/09)
With 1μg of DNA, we manage to obtain colonies with ECE112. We will check our transformation with starch plates.

Check ECE112 stock
- Single and double digest with XbaI and EcoRI



- Gel
 * Lane 3 : HyperladderI
 * Lane 4 : ECE112, 100mL flask, single digest with XbaI (expected size about 10,000bp)
 * Lane 5 : ECE112, 100mL flask, single digest with EcoRI (expected size about 10,000bp)
 * Lane 6 : ECE112, 100mL flask, double digest (expected size about 3200bp and 6,900bp)
 * Lane 7 : ECE112, 10mL flask, single digest with XbaI (expected size about 10,000bp)
 * Lane 8 : ECE112, 10mL flask, single digest with EcoRI (expected size about 10,000bp)
 * Lane 9 : ECE112, 10mL flask, double digest (expected size about 3200bp and 6,900bp)
 * Lane 10 : HyperladderI

- Results : ok

Starch plates
- Use blank plates

- Melt 100mL of Soft Agar, and add slowly 1g of starch

- Mix, boil to sterilize

- Pour on blank plate

- Put 4 or 5 colonies on each plate


 * IA751 from blank plate (positive control)
 * IA771 from blank plate (negative control)
 * IA751 transformed with ECE112 (Cm5, 29/08)
 * IA751 control (Cm5, 29/08)
 * IA751 + ECE112 (Cm5, 02/09)
 * IA751 + ECE153 (Cm5, 02/09)
 * IA751 + ECE153 (Cm5, 29/08)