IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/07/28: Difference between revisions

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{{Cambridge08a}}
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|style="background-color: #EEE"|[[Image:IgemCam.jpg|200px]]<span style="font-size:22px;"> Turing Pattern Formation</span>
|style="background-color: #444444"|[[Image:Signalling_button.gif|200px|center]]
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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__NOTOC__
==Results from previous days==
==Results from previous days==
* Result of the gel
* '''Result of the gel'''
 
* Lane3 : hyperladder1
* Lane4 : I746101
* Lane5 : I746001
 
[[Image:photo1.gif|300px|center]]


We wanted to check the size of our biobricks.  
We wanted to check the size of our biobricks.  
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So, the size of our biobricks are ok.  
So, the size of our biobricks are ok.  
* Results of antibiotic plates from yesterday
For Cm, 35μg/mL should be enough to kill B.S.
For Amp,




==Wet Work==
==Wet Work==


* Extract plasmid ppL82
- Digest
 
We had 2 samples of ppL82 in DH5α in LB solution, one from the frozen glycerol tube and one from a colony picked on a plate.
Normally plasmid and cells should not be kept in freezer. So, we want to extract this plasmid, check its size and then keep it in the fridge.
We do that for the 2 different samples.
 
- Plasmid Miniprep
 
 
 
 
* Preparation of different stocks of strains and plasmids
 
 
'''PNZ8901'''
 
- Plate a single colony (from 23/07/08) on a Cm plate
 
- Incubate at 37°C
 
 
- Grow the entire frozen glycerol tube in 20mL of LB without antibiotic to check if this stock is still good (we will check the plasmid on a gel tomorrow)
 
- Incubate at 37°C
 
 
'''MC1061'''
 
-Pick e single colony of MC1061 (from 22/07/08) and put it in 10mL of LB (to make glycerol stocks)


-Incubate at 37°C
# For biobricks I746001 and I746101, use digest enzymes EcoRI and SpeI, and 2μL of DNA
#: Add 15μL of SDW, 2μL of 10X Fast Digest Buffer, 2μL of DNA, 1μL of EcoRI and 1μL of SpeI
# For ppL82 (from plate and tube), use digest enzymes BamHI and PstI and add 15μL of DNA
#: Add 2μL of SDW, 2μL of 10X Fast Digest Buffer, 15μL of DNA, 1μL of BamHI and 1μL of PstI
# Microfuge tubes
# Incubate 10 min at 37°C
# Heat shock for 5min at 80°C


- Gel : add 4μL of dye and 21μL of samples


'''1A1'''
- Add 100μL of LB+1A (mix of friday) and 10mL of LB
- Incubate at 37°C
==Prepare Medium A==
- Add 81mL of Sterile Water, 10mL of 10X Medium A base and 9mL of 10X Bacillus salts


* '''New plates'''


-Plate I746001 and I746101 on Kn plates from Amp plates from 24/07/2008.


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__NOTOC__

Revision as of 09:25, 5 September 2008

 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Results from previous days

  • Result of the gel
  • Lane3 : hyperladder1
  • Lane4 : I746101
  • Lane5 : I746001

We wanted to check the size of our biobricks. For I746101, we have a band of about 3000kb, which is the size of the vectorm and a band of about 2000kb. The size of the biobrick is 2057kp, so it should be ok.

For I746001, we hqve the same band of 3000kb (the vector) and a band of about 1000kb. The size of the biobrick is 896kb.

So, the size of our biobricks are ok.


Wet Work

- Digest

  1. For biobricks I746001 and I746101, use digest enzymes EcoRI and SpeI, and 2μL of DNA
    Add 15μL of SDW, 2μL of 10X Fast Digest Buffer, 2μL of DNA, 1μL of EcoRI and 1μL of SpeI
  2. For ppL82 (from plate and tube), use digest enzymes BamHI and PstI and add 15μL of DNA
    Add 2μL of SDW, 2μL of 10X Fast Digest Buffer, 15μL of DNA, 1μL of BamHI and 1μL of PstI
  3. Microfuge tubes
  4. Incubate 10 min at 37°C
  5. Heat shock for 5min at 80°C

- Gel : add 4μL of dye and 21μL of samples


  • New plates

-Plate I746001 and I746101 on Kn plates from Amp plates from 24/07/2008.

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