IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/07/23: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
(5 intermediate revisions by 3 users not shown)
Line 1: Line 1:
<hr width="810px">
<div id="about">
{{Cambridge08a}}
</div>
<hr width="810px">
{|{{table}} width="800"
{|{{table}} width="800"
|-
|-
|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>}}
|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>}}
|-
|-
| colspan="2"|
| colspan="2"|
==Wet work==
* Take one colony from each of the plates grown from the tubes yesterday and plated them again on LA without antibiotics
'''1/ Purpose''' Check antibbiotic resistance of our different strains
{|class="wikitable" style="text-align:center" border="1"
|-
! Strain
! Plasmid
! Extracted from
! Antibiotic use
! Concentration of antibiotic
|-
| DH5alpha || PP182 || tube || Amp || 100
|-
| MC1061 || NONE || tube || Cm || 35
|-
| MC1061 || PNZ8901 || tube || Cm || 35
|-
| DH5alpha || PP182 || plate || Amp || 100
|-
| MC1061 || NONE || plate || Cm || 35
|-
| MC1061 || PNZ8901 || plate || Cm || 35
|-
|}
'''2/ Purpose''' : Find the right concentration of antibiotic so that B. subtilis survive
- Grow 15uL of B. 1A1 (frozen tube) in 5mL of LB
- Incubate at 37°C
'''3/ Purpose ''': Grow plasmids in TOP10, transformation
4 plasmids :
* I746000
* I746100
* I746101
* I746001
1 control : PUC19
- Add 20uL of TOP10 competent cells and 0.5uL of Plasmid in an eppendorf
- 30min on ice
- Heat shock : 60s at 42°C


- 2min on ice
==Dry work==
* found assay to test for AIP
* found shuttle vector for constitutive expression of GFP
* found protocol for Bacillus subtilis transformation
** Unsolved question: do we need to use Tryptone Blood Agar base? can we use regular L broth/agar?


- Add 89.5uL of SOC


- 60min at 37°C
- Put the mix on plate with ampicillin resistance
- Incubate at 37°C




Line 71: Line 24:
<!-- ## Do not edit below this line unless you know what you are doing. ## -->
<!-- ## Do not edit below this line unless you know what you are doing. ## -->
|}
|}
__NOTOC__

Revision as of 09:23, 5 September 2008

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

Dry work

  • found assay to test for AIP
  • found shuttle vector for constitutive expression of GFP
  • found protocol for Bacillus subtilis transformation
    • Unsolved question: do we need to use Tryptone Blood Agar base? can we use regular L broth/agar?



Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:Cambridge/2008/Notebook/Turing_Pattern_Formation/2008/07/23&oldid=238720"