IGEM:Cambridge/2008/Notebook/Bacillus/2008/08/22: Difference between revisions

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(Autocreate 2008/08/22 Entry for IGEM:Cambridge/2008/Notebook/Bacillus)
 
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==Entry title==
==Stock ECE112, 166, 153==
* Insert your content here.


- Plasmid miniprep (2 tubes of ECE112, 2 tubes of ECE166 and 1 tube of ECE153)


- Nanodrop


<!-- ## Do not edit below this line unless you know what you are doing. ## -->
==Transformation of Bacillus==
 
* For strain IA751 : transform with ECE166, 171, 153 and 112
 
* For strain IA771 : transform with ECE166, 171
 
* for glycerol stocks (one with 100μL of medium B and glycerol, and one with only glycerol) : transform with ECE166
 
 
- We used the same protocol than before with a few changes :
 
* add sterile 50% glucose in medium A
* don't keep samples after using them for spectrophotometer
* spin each tubes and keep only 100μL of medium A (to concentrate cells) before adding DNA
 
{|class="wikitable" style="text-align:center" border="1"
|-
! Time (min) !! 0 !! 20 !! 40 !! 60 !! 80 !! 100 !! 120 !! 130 !! 140
|-
| OD650 for IA751 || 0.1817 || 0.1870 || 0.1929 || 0.2115 || 0.2543 || 0.3291 || 0.3695 || 0.4188 ||0.4224
|-
| OD650 for IA751 ||0.1818 || 0.1874 || 0.1907 || 0.2088 || 0.2452 || 0.3182 || 0.3575 || 0.4228 || 0.4314
|-
|}
|}


[[Image:graph.gif|500px|center]]


__NOTOC__
- For glycerol stocks, resuspend cells into 100μL of medium B, add DNA and wait for 30min, end plate them

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Stock ECE112, 166, 153

- Plasmid miniprep (2 tubes of ECE112, 2 tubes of ECE166 and 1 tube of ECE153)

- Nanodrop

Transformation of Bacillus

  • For strain IA751 : transform with ECE166, 171, 153 and 112
  • For strain IA771 : transform with ECE166, 171
  • for glycerol stocks (one with 100μL of medium B and glycerol, and one with only glycerol) : transform with ECE166


- We used the same protocol than before with a few changes :

  • add sterile 50% glucose in medium A
  • don't keep samples after using them for spectrophotometer
  • spin each tubes and keep only 100μL of medium A (to concentrate cells) before adding DNA
Time (min) 0 20 40 60 80 100 120 130 140
OD650 for IA751 0.1817 0.1870 0.1929 0.2115 0.2543 0.3291 0.3695 0.4188 0.4224
OD650 for IA751 0.1818 0.1874 0.1907 0.2088 0.2452 0.3182 0.3575 0.4228 0.4314

- For glycerol stocks, resuspend cells into 100μL of medium B, add DNA and wait for 30min, end plate them

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