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| Line 17: |
Line 17: |
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| - Add 5μL of ligation products, and 1.2μL of PUC9 | | - Add 5μL of ligation products, and 1.2μL of PUC9 |
|
| |
| ==Xylose test==
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| * Preparation of xylose tube
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|
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| - Add 1mL of culture from yesterday, 1mL of xylose (0.1g/mL) and 8mL of LB and 45μL of Spc (do that for the 4 different colonies)
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|
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| - Incubate 5hours
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| - Observation with microscope : some green task, but they do not really match with bacillus cells, not conclusive!
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|
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| ==Starch plate==
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| - Add 5mL of iodine solution on starch plate, wait for 1min, throw the liquid
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|
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| - Result
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| {|class="wikitable" style="text-align:center" border="1"
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| |-
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| ! Strain !! added vector !! Observation !! Conclusion
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| |-
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| | IA771 || none || no clearance zones || ok
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| |-
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| | IA751 || none || big clearance zones || ok
| |
| |-
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| | IA751 || ECE153 || no clearance zones || trnsformation worked!
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| |-
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| | IA751 or IA771 || none or ECE171 || big clearance zones || we have some colonues on control plate with IA751, contamination? or antibiotic resistance?
| |
| |-
| |
| |}
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|
| |
| ==Ery test==
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| - IA771 alive and IA751 dead, confirmation that we did not mix plates (same conclusion than with starch plate, last line)
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|
| |
| ==Test glycerol stocks of competent bacillus==
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|
| |
| - Pellet bacillus cells, add 100μL of prewarmed mediumB, add 0.6μg of DNA, incubate 30min at 37°C by shaking, plate
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|
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| {|class="wikitable" style="text-align:center" border="1"
| |
| |-
| |
| ! Strain !! added vector !! Antibiotic
| |
| |-
| |
| | IA771 || none || blank plate (to check they are alive)
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| |-
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| | IA771 || none || Cm5 (negative control)
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| |-
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| | IA771 || ECE166 || Cm5
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| |-
| |
| | IA751 || none || blank plate (to check they are alive)
| |
| |-
| |
| | IA751 || none || Cm5 (negative control)
| |
| |-
| |
| | IA751 || none || Spc50 (negative control)
| |
| |-
| |
| | IA751 || ECE112 || Cm5
| |
| |-
| |
| | IA751 || ECE153 || Spc50
| |
| |}
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| <!-- ## Do not edit below this line unless you know what you are doing. ## -->
| |
| |}
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| __NOTOC__
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