IGEM:IMPERIAL/2006/project/Oscillator/project browser/Predator Construct/Implementation: Difference between revisions

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==Ligation strategy==
==Ligation strategy==  
*For predator sensing characterisation, we planned to build :
 
**[http://parts.mit.edu/registry/index.php/Part:BBa_J37016 J37016]
<u><font size="2">'''Ligating the Polycystronic Predator Construct:'''</font></u>
**[http://parts.mit.edu/registry/index.php/Part:BBa_J37020 J37020]
__NOTOC__
*For predator killing characterisation, we planned to build :
{| border="1" cellpadding="0" width="100%"
**[http://parts.mit.edu/registry/index.php/Part:BBa_J37023 J37023]
|-
**[http://parts.mit.edu/registry/index.php/Part:BBa_J37024 J37024]
!style="background:#E6B3E0" height="25pt" width="80pt" |For !! style="background:#E6B3E0" |Stage 1 !! style="background:#E6B3E0" |Stage 2 !! style="background:#E6B3E0" |Stage 3 !! style="background:#E6B3E0" |Stage 4
*For overall predator construct, we planned to build
|-
**[http://parts.mit.edu/registry/index.php/Part:BBa_J37035 J37035]
!style="background:#D9F2ED"|<center><bbpart>J37023</bbpart><br>(<bbpart>"PCR'd"</bbpart>)</center> !!style="background:#CCEBAD" | <center>[[Image:J37023 logo.png]]</center> || ||  ||
**[http://parts.mit.edu/registry/index.php/Part:BBa_J37036 J37036]
|-
!style="background:#D9F2ED"|<center><bbpart>J37024</bbpart>=<br>(<bbpart>B0034</bbpart>+<bbpart>J37023</bbpart>)</center>|| !!style="background:#CCEBAD" | <center>[[Image:B0034 logo.png]][[Image:Plus sign.png|20px]][[Image:J37023 logo.png]]</center> || ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37025</bbpart>=<br>(<bbpart>J37024</bbpart>+<bbpart>B0014</bbpart>)</center>||  ||  !!style="background:#CCEBAD" | <center>[[Image:B0034 logo.png]][[Image:J37024 logo.png]]</center> ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37033</bbpart>=<br>(<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)</center> !!style="background:#CCEBAD" | <center>[[Image:B0034 logo.png]][[Image:Plus sign.png|20px]][[Image:C0062 logo.png]]</center> || || ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37019</bbpart>=<br>(<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)</center>|| !!style="background:#CCEBAD" | <center>[[Image:R0062 logo.png]][[Image:Plus sign.png|20px]][[Image:B0034 logo.png]][[Image:C0062 logo.png]]</center> || ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37036</bbpart>=<br>(<bbpart>J37019</bbpart>+<bbpart>J37025</bbpart>)</center>|| ||  || !!style="background:#CCEBAD" | <center>[[Image:J37036 logo.png]]</center>
|}
 
 
 
<u><font size="2">'''Ligating the Monocystronic Predator Construct:'''</font></u>
__NOTOC__
{| border="1" cellpadding="0" width="100%"
|-
!style="background:#E6B3E0" height="25pt" width="80pt" |For !! style="background:#E6B3E0" |Stage 1 !! style="background:#E6B3E0" |Stage 2 !! style="background:#E6B3E0" |Stage 3 !! style="background:#E6B3E0" |Stage 4 !! style="background:#E6B3E0" |Stage 5
|-
!style="background:#D9F2ED"|<center><bbpart>J37023</bbpart><br>(<bbpart>"PCR'd"</bbpart>)</center> !!style="background:#CCEBAD" | <center>[[Image:J37023 logo.png|30px]]</center> || || || ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37024</bbpart>=<br>(<bbpart>J37023</bbpart>+<bbpart>B0014</bbpart>)</center>|| !!style="background:#CCEBAD" | <center>[[Image:J37023 logo.png|30px]][[Image:Plus sign.png|20px]][[Image:J37024 logo.png|70px]]</center> || || ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37025</bbpart>=<br>(<bbpart>B0034</bbpart>+<bbpart>J37024</bbpart>)</center>||  ||  !!style="background:#CCEBAD" | <center>[[Image:J37023 logo.png|30px]][[Image:J37024 logo.png|70px]]</center> || ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37033</bbpart>=<br>(<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)</center> !!style="background:#CCEBAD" | <center>[[Image:B0034 logo.png|30px]][[Image:Plus sign.png|20px]][[Image:C0062 logo.png|30px]]</center> || || || ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37019</bbpart>=<br>(<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)</center>|| !!style="background:#CCEBAD" | <center>[[Image:R0062 logo.png|30px]][[Image:Plus sign.png|20px]][[Image:B0034 logo.png|30px]][[Image:C0062 logo.png|30px]]</center> || || ||
|-
!style="background:#D9F2ED"|<center><bbpart>J37031</bbpart>=<br>(<bbpart>J37019</bbpart>+<bbpart>B0014</bbpart>)</center>|| || !!style="background:#CCEBAD" |[[Image:R0062 logo.png|30px]][[Image:B0034 logo.png|30px]][[Image:C0062 logo.png|30px]][[Image:Plus sign.png|20px]][[Image:B0014 logo.png|50px]]
|-
!style="background:#D9F2ED"|<center><bbpart>J37018</bbpart>=<br>(<bbpart>J37031</bbpart>+<bbpart>R0062</bbpart>)</center>||  ||  || !!style="background:#CCEBAD" | <center>[[Image:R0062 logo.png|30px]][[Image:B0034 logo.png|30px]][[Image:C0062 logo.png|30px]][[Image:B0014 logo.png|50px]][[Image:Plus sign.png|20px]][[Image:R0062 logo.png|30px]]</center>
|-
!style="background:#D9F2ED"|<center><bbpart>J37035</bbpart>=<br>(<bbpart>J37018</bbpart>+<bbpart>J37025</bbpart>)</center>|| || || || !!style="background:#CCEBAD" | <center>[[Image:R0062 logo.png|30px]][[Image:B0034 logo.png|30px]][[Image:C0062 logo.png|30px]][[Image:B0014 logo.png|50px]][[Image:R0062 logo.png|30px]][[Image:J37023 logo.png|30px]][[Image:J37024 logo.png|70px]]</center>
|}
 
*More information on ligations can be found on the [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/Ligation_Page Ligation Page].


==Quality control==
==Quality control==
*define the quality control used at each step
*define possible enzymes to be used and their outcomes
*define design of primers to be used in sequencing
__NOTOC__
__NOTOC__
{| border="1" cellpadding="0" width="100%"
{| border="1" cellpadding="0" width="100%"
|-
|-
!height="25pt" width="80pt"| Part !! Built !! Sequencing Results !! Primers Used for Sequencing
!height="25pt" width="80pt"| Part !! Logo !! Built !! Sequenced !! Primers Used for Sequencing
|-
|-
|Put part logo and name here || has it been built? || sequencing analysis and link to sequence results || primers used
| <center><bbpart>J37018</bbpart></center> || [[Image:J37018 logo.png]] || No || N/A || N/A
|-
|-
|Put part logo and name here || has it been built? || sequencing analysis and link to sequence results || primers used
| <center><bbpart>J37019</bbpart></center> || [[Image:J37019 part.png]] || Yes || [http://openwetware.org/images/e/e3/J37019_sequencing.doc Yes] || [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/parts/Primers Primers used]
|-
|-
| <center><bbpart>J37023</bbpart></center> || [[Image:J37023 logo.png]] || Yes || [http://openwetware.org/images/b/b3/J37025sequencing.doc Yes] <br> Note: This part was sequenced as a part of J37025 || [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/parts/Primers Primers used]
|-
| <center><bbpart>J37024</bbpart></center> || [[Image:J37024 logo.png]] || No || N/A || N/A
|-
| <center><bbpart>J37025</bbpart></center> || [[Image:J37025 logo.png]] || Yes || [http://openwetware.org/images/b/b3/J37025sequencing.doc Yes] || [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/parts/Primers Primers used]
|-
| <center><bbpart>J37031</bbpart></center> || [[Image:J37031 logo.png]] || Yes || No || N/A
|-
| <center><bbpart>J37033</bbpart></center> || [[Image:J37033 logo.png]] || Yes || No || N/A
|-
| <center><bbpart>J37035</bbpart></center> || [[Image:J37035 logo.png]] || No || N/A || N/A
|-
| <center><bbpart>J37036</bbpart></center> || [[Image:J37036 part.png]] || No || N/A || N/A
|}
|}


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*the final assembly was not successful
*the final assembly was not successful
*The AiiA gene available from the registry seemed non-functional. We sourced new AiiA to which we then PCR'd LVA-tag and immunotag. See [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/Oscillator/project_browser/Test_Killing_Predator_Construct/Design design section for this part] for more information]
*Reasons for parts not being built include time constraint (and perhaps non-functional registry parts).
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Latest revision as of 09:43, 30 October 2006

Super Parts Molecular Prey-Predator Oscillator
Actual Part
Sub Parts Test Sensing Predator Construct Test Killing Predator Construct


Ligation strategy

Ligating the Polycystronic Predator Construct:

For Stage 1 Stage 2 Stage 3 Stage 4
<bbpart>J37023</bbpart>
(<bbpart>"PCR'd"</bbpart>)
<bbpart>J37024</bbpart>=
(<bbpart>B0034</bbpart>+<bbpart>J37023</bbpart>)
<bbpart>J37025</bbpart>=
(<bbpart>J37024</bbpart>+<bbpart>B0014</bbpart>)
<bbpart>J37033</bbpart>=
(<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)
<bbpart>J37019</bbpart>=
(<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)
<bbpart>J37036</bbpart>=
(<bbpart>J37019</bbpart>+<bbpart>J37025</bbpart>)


Ligating the Monocystronic Predator Construct:

For Stage 1 Stage 2 Stage 3 Stage 4 Stage 5
<bbpart>J37023</bbpart>
(<bbpart>"PCR'd"</bbpart>)
<bbpart>J37024</bbpart>=
(<bbpart>J37023</bbpart>+<bbpart>B0014</bbpart>)
<bbpart>J37025</bbpart>=
(<bbpart>B0034</bbpart>+<bbpart>J37024</bbpart>)
<bbpart>J37033</bbpart>=
(<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)
<bbpart>J37019</bbpart>=
(<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)
<bbpart>J37031</bbpart>=
(<bbpart>J37019</bbpart>+<bbpart>B0014</bbpart>)
<bbpart>J37018</bbpart>=
(<bbpart>J37031</bbpart>+<bbpart>R0062</bbpart>)
<bbpart>J37035</bbpart>=
(<bbpart>J37018</bbpart>+<bbpart>J37025</bbpart>)

Quality control

Part Logo Built Sequenced Primers Used for Sequencing
<bbpart>J37018</bbpart>
 No N/A N/A
<bbpart>J37019</bbpart>
 Yes Yes Primers used
<bbpart>J37023</bbpart>
 Yes Yes
Note: This part was sequenced as a part of J37025		 Primers used
<bbpart>J37024</bbpart>
 No N/A N/A
<bbpart>J37025</bbpart>
 Yes Yes Primers used
<bbpart>J37031</bbpart>
 Yes No N/A
<bbpart>J37033</bbpart>
 Yes No N/A
<bbpart>J37035</bbpart>
 No N/A N/A
<bbpart>J37036</bbpart>
 No N/A N/A

WetLab work entries

  • the final assembly was not successful
  • The AiiA gene available from the registry seemed non-functional. We sourced new AiiA to which we then PCR'd LVA-tag and immunotag. See design section for this part for more information]
  • Reasons for parts not being built include time constraint (and perhaps non-functional registry parts).

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