IGEM:Cambridge/2008/Notebook/Bacillus/2008/09/05: Difference between revisions

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==Transformation of Bacillus==


* Result of Nanodrop
{|class="wikitable" style="text-align:center" border="1"
|-
!  Vector !! 260/280 !! ng/μL
|-
| ECE112 || 1.75 || 64.6
|-
| ECE166 || 172 || 138.6
|}
- Prepare medium A with tryptophan, and medium B
- add 5mL of medium A in 3 different tubes, inoculate each tube with colonies (1A1, IA751 ans IA771)
- Check OD every 20min
- Incubate 90min
- Add 0.45mL of medium B and 0.05mL of culture in Ependorf tubes
- Incubate 90min
- Transform : add 1μg of DNA (some with ECE112, some with ECE166) (so you need to nanodrop samples before!)
- Incubate 30min
- Pipette 200μL of solution, spread it on aech plate, wait 10min, and do it again
- Incubate 24hours
- A few glycerol tubes to stock cells : add 2/7 glycerol to cell tubes
- Transformation from glycerol stock from 30/07/2008
- Spin glycerol stocks, pipette out glycerol
- Add 0.5mL of medium B, incubate for 1 hour
- Add 10μL of ECE112 (640ng)
- Incubate 2hours
- Plate 200μL, and 10min later, still 200μL
==New stocks==
- Do glycerol stock of I746001 and I746101 (no sterile glycerol)
- Put IA751, IA771 in 10mL LB
- Put ECE 176 in 10mL LB + antibiotic
- Reinoculate the tube of LB from yesterday with ECE166 plate
- ECE176 replated onto Amp100 + Cm5


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Revision as of 04:58, 5 September 2008

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