Difference between revisions of "Status Of Building Parts"

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MWSU + Davidson Status of Building Sender and Receiver
 
MWSU + Davidson Status of Building Sender and Receiver
 +
 +
[http://partsregistry.org/AHL '''List of auto-inducers and their Sigma catalog numbers'''.]
  
 
plux-lac hybrid promoter:
 
plux-lac hybrid promoter:
 
* [http://partsregistry.org/Part:BBa_I751502 I751502]
 
* [http://partsregistry.org/Part:BBa_I751502 I751502]
 
* In the process of building another plux-lac hybrid promoter that is repressed by HSL and lacI
 
* In the process of building another plux-lac hybrid promoter that is repressed by HSL and lacI
−
Responsible: Robert Cool, Erin Feeney and Pallavi Penumetcha
+
Responsible: Robert Cool, Erin Feeney and Pallavi Penumetcha <br>
 +
<br>
 +
'''Parts we have Mini-Preps and Digestions for'''
 +
{| border="1" cellpadding="2"
 +
!width="50"|Part
 +
!width="225"|Description
 +
!width="65"|Size of Insert (w/ BioBrick ends)
 +
!width="150"|Verification using Gel Electrophoresis
 +
!width="45"|Glycerol Stock
 +
|-
 +
|BBa_F2620|| Accepts 3OC6HSL as an input and outputs PoPS||1102 bp||Appears to be reversed with J04430|| Yes
 +
|-
 +
|BBa_J04430|| GFP generator switched on by IPTG||1124 bp|| Appears to be reversed with F2620|| Yes
 +
|-
 +
|BBa_P0440|| Produces tetR||881 bp|| Verified|| Yes
 +
|-
 +
|BBa_E0240|| GFP generator|| 917 bp||Verified|| Yes
 +
|-
 +
|BBa_I132623|| Lux receiver that produces luxR and outputs EYFP|| 2085 bp|| Much smaller on gel ||Yes
 +
|-
 +
|BBa_I0426|| Produces lasR and outputs ECFP followed by a lasR-PAI regulated promoter that outputs EYFP ||2839 bp|| Verified|| Yes
 +
|-
 +
|BBa_C0061|| Produces luxI (w/ LVA tag) ||659 bp|| Verified|| Yes
 +
|-
 +
|BBa_C0261|| Produces luxI (w/ RBS)|| 702 bp|| No successful Mini-Preps
 +
|-
 +
|BBa_R0062|| luxR and HSL regulated promoter|| 96 bp|| Verified|| Yes
 +
|-
 +
|BBa_I0466|| RhlR protein generator|| 983 bp|| Verified|| Yes
 +
|-
 +
|BBa_I0407|| Produces lasI and outputs ECFP|| 1674 bp|| Verified|| Yes
 +
|-
 +
|BBa_R0079|| LasR and PAI regulated promoter || 198 bp|| Verified|| Yes
 +
|-
 +
|BBa_R0063|| LuxR and HSL regulated promoter|| 192 bp|| No successful Mini-Preps
 +
|-
 +
|BBa_K09100|| Receiver for AHL and outputs GFP|| 1987 bp|| Verified|| Yes
 +
|-
 +
|BBa_J04450|| RFP coding device switched on by IPTG|| 2110 bp|| Much smaller on gel
 +
|-
 +
|BBa_C0079|| lasR activator|| 797|| Verified|| Yes
 +
|-
 +
|BBa_S03156|| Ribosomal binding site followed by lasr coding region|| 840|| Verified|| Yes
 +
|-
 +
|BBa_S03154|| Ribosomal binding site followed by lasI coding region|| 726||
 +
|-
 +
|BBa_T9002||Inputs C06HSL and outputs GFP|| 1986||
 +
|}
 +
-We will be sending all of these parts to be sequenced <br>
 +
<br>
  
 +
'''Gels used to verify parts''' <br>
 +
''Gel A'' <br>
 +
These are the parts run on a 0.5% gel.<br>
 +
[[Image:gelA.jpg]]<br>
 +
'''Top portion of Gel A''' <br>
 +
{| border="1" cellpadding="2"
 +
!width="50"|Lane
 +
!width="50"|Part
 +
|-
 +
|1|| MW
 +
|-
 +
|2|| F0260
 +
|-
 +
|3|| J04430
 +
|-
 +
|4||P0440
 +
|-
 +
|5|| E0240
 +
|-
 +
|6|| I13263
 +
|-
 +
|7|| J0440
 +
|-
 +
|} <br>
 +
'''Bottom portion of Gel A''' <br>
 +
{| border="1" cellpadding="2"
 +
!width="50"|Lane
 +
!width="50"|Part
 +
|-
 +
|1|| MW
 +
|-
 +
|2|| C0061
 +
|-
 +
|3|| I0466
 +
|-
 +
|4|| I0407
 +
|-
 +
|5|| I0426
 +
|-
 +
|6|| K09100
 +
|-
 +
|} <br>
 +
GelB<br>
 +
These parts were run on a 2.5% gel.<br>
 +
[[Image:gelB.jpg]]<br>
 +
{| border="1" cellpadding="2"
 +
!width="50"|Lane
 +
!width="50"|Part
 +
|-
 +
|1|| MW
 +
|-
 +
|2|| R0062
 +
|-
 +
|3|| R0079
 +
|-
 +
|9|| pTet-lac PCR product
 +
|-
 +
|10|| MW
 +
|-
 +
|} <br>
 
[http://parts.mit.edu/igem07/index.php/USTC '''Look at USTC 2007 for logic gates''']
 
[http://parts.mit.edu/igem07/index.php/USTC '''Look at USTC 2007 for logic gates''']
  
Line 17: Line 128:
  
 
[http://partsregistry.org/Part:BBa_S03632 '''S03632'''] (ligations needed)<br>
 
[http://partsregistry.org/Part:BBa_S03632 '''S03632'''] (ligations needed)<br>
−
[[Image:S03632.jpg]]  
+
[[Image:S03632.jpg]] <br>
−
 
+
'''But BBa_S03632 is not available, so we will need to build our own version using the 3 parts below:'''<br>
−
Parts: [http://partsregistry.org/Part:BBa_R0011 '''R0011''' ][http://partsregistry.org/wiki/index.php?title=Part:BBa_C0261 '''C0261'''] [http://partsregistry.org/Part:BBa_B0015 '''B0015''']
+
Parts: [http://partsregistry.org/Part:BBa_R0011 '''R0011''' ] + [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0261 '''C0261'''] + [http://partsregistry.org/Part:BBa_B0015 '''B0015''']<br>
 +
[[Image:R0011b.jpg]] + [[Image:C0261.jpg]] + [[Image:B0015.jpg]]
  
 
----
 
----

Latest revision as of 14:14, 18 June 2008

MWSU + Davidson Status of Building Sender and Receiver

List of auto-inducers and their Sigma catalog numbers.

plux-lac hybrid promoter:

  • I751502
  • In the process of building another plux-lac hybrid promoter that is repressed by HSL and lacI

Responsible: Robert Cool, Erin Feeney and Pallavi Penumetcha

Parts we have Mini-Preps and Digestions for

Part Description Size of Insert (w/ BioBrick ends) Verification using Gel Electrophoresis Glycerol Stock
BBa_F2620 Accepts 3OC6HSL as an input and outputs PoPS 1102 bp Appears to be reversed with J04430 Yes
BBa_J04430 GFP generator switched on by IPTG 1124 bp Appears to be reversed with F2620 Yes
BBa_P0440 Produces tetR 881 bp Verified Yes
BBa_E0240 GFP generator 917 bp Verified Yes
BBa_I132623 Lux receiver that produces luxR and outputs EYFP 2085 bp Much smaller on gel Yes
BBa_I0426 Produces lasR and outputs ECFP followed by a lasR-PAI regulated promoter that outputs EYFP 2839 bp Verified Yes
BBa_C0061 Produces luxI (w/ LVA tag) 659 bp Verified Yes
BBa_C0261 Produces luxI (w/ RBS) 702 bp No successful Mini-Preps
BBa_R0062 luxR and HSL regulated promoter 96 bp Verified Yes
BBa_I0466 RhlR protein generator 983 bp Verified Yes
BBa_I0407 Produces lasI and outputs ECFP 1674 bp Verified Yes
BBa_R0079 LasR and PAI regulated promoter 198 bp Verified Yes
BBa_R0063 LuxR and HSL regulated promoter 192 bp No successful Mini-Preps
BBa_K09100 Receiver for AHL and outputs GFP 1987 bp Verified Yes
BBa_J04450 RFP coding device switched on by IPTG 2110 bp Much smaller on gel
BBa_C0079 lasR activator 797 Verified Yes
BBa_S03156 Ribosomal binding site followed by lasr coding region 840 Verified Yes
BBa_S03154 Ribosomal binding site followed by lasI coding region 726
BBa_T9002 Inputs C06HSL and outputs GFP 1986

-We will be sending all of these parts to be sequenced

Gels used to verify parts
Gel A
These are the parts run on a 0.5% gel.
GelA.jpg
Top portion of Gel A

Lane Part
1 MW
2 F0260
3 J04430
4 P0440
5 E0240
6 I13263
7 J0440

Bottom portion of Gel A

Lane Part
1 MW
2 C0061
3 I0466
4 I0407
5 I0426
6 K09100

GelB
These parts were run on a 2.5% gel.
GelB.jpg

Lane Part
1 MW
2 R0062
3 R0079
9 pTet-lac PCR product
10 MW

Look at USTC 2007 for logic gates

T9002 - no colonies
T9002.jpg

F2622 + E0240 (ligated/colonies coming up)
F2622.jpg + E0240.jpg

S03632 (ligations needed)
S03632.jpg
But BBa_S03632 is not available, so we will need to build our own version using the 3 parts below:
Parts: R0011 + C0261 + B0015
R0011b.jpg + C0261.jpg + B0015.jpg

R0011 + F1610 (minis, frag needed/colonies coming up, minis thurs)
R0011b.jpg + F1610.jpg

F2620 + E0240 (minis growing/ frag available)
F2620b.jpg + E0240.jpg

P0440
P0440.jpg

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