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: | ||
| Line 12: | Line 14: | ||
!width="65"|Size of Insert (w/ BioBrick ends) | !width="65"|Size of Insert (w/ BioBrick ends) | ||
!width="150"|Verification using Gel Electrophoresis | !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 | + | |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 | + | |BBa_J04430|| GFP generator switched on by IPTG||1124 bp|| Appears to be reversed with F2620|| Yes |
|- | |- | ||
| − | |BBa_P0440|| Produces tetR||881 bp|| Verified | + | |BBa_P0440|| Produces tetR||881 bp|| Verified|| Yes |
|- | |- | ||
| − | |BBa_E0240|| GFP generator|| 917 bp||Verified | + | |BBa_E0240|| GFP generator|| 917 bp||Verified|| Yes |
|- | |- | ||
| − | |BBa_I132623|| Lux receiver that produces luxR and outputs EYFP|| 2085 bp|| Much smaller on gel | + | |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 | + | |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 | + | |BBa_C0061|| Produces luxI (w/ LVA tag) ||659 bp|| Verified|| Yes |
|- | |- | ||
|BBa_C0261|| Produces luxI (w/ RBS)|| 702 bp|| No successful Mini-Preps | |BBa_C0261|| Produces luxI (w/ RBS)|| 702 bp|| No successful Mini-Preps | ||
|- | |- | ||
| − | |BBa_R0062|| luxR and HSL regulated promoter|| 96 bp|| Verified | + | |BBa_R0062|| luxR and HSL regulated promoter|| 96 bp|| Verified|| Yes |
|- | |- | ||
| − | |BBa_I0466|| RhlR protein generator|| 983 bp|| Verified | + | |BBa_I0466|| RhlR protein generator|| 983 bp|| Verified|| Yes |
|- | |- | ||
| − | |BBa_I0407|| Produces lasI and outputs ECFP|| 1674 bp|| Verified | + | |BBa_I0407|| Produces lasI and outputs ECFP|| 1674 bp|| Verified|| Yes |
|- | |- | ||
| − | |BBa_R0079|| LasR and PAI regulated promoter || 198 bp|| Verified | + | |BBa_R0079|| LasR and PAI regulated promoter || 198 bp|| Verified|| Yes |
|- | |- | ||
| − | |BBa_R0063|| LuxR and HSL regulated promoter|| 192 bp|| | + | |BBa_R0063|| LuxR and HSL regulated promoter|| 192 bp|| No successful Mini-Preps |
|- | |- | ||
| − | |BBa_K09100|| Receiver for AHL and outputs GFP|| 1987 bp|| Verified | + | |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_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> | -We will be sending all of these parts to be sequenced <br> | ||
<br> | <br> | ||
| + | |||
'''Gels used to verify parts''' <br> | '''Gels used to verify parts''' <br> | ||
''Gel A'' <br> | ''Gel A'' <br> | ||
| Line 88: | Line 99: | ||
|- | |- | ||
|} <br> | |} <br> | ||
| − | + | GelB<br> | |
| − | These parts were run on a 2.5% gel. <br> | + | These parts were run on a 2.5% gel.<br> |
[[Image:gelB.jpg]]<br> | [[Image:gelB.jpg]]<br> | ||
{| border="1" cellpadding="2" | {| border="1" cellpadding="2" | ||
| Line 103: | Line 114: | ||
|9|| pTet-lac PCR product | |9|| pTet-lac PCR product | ||
|- | |- | ||
| − | |10|| | + | |10|| MW |
|- | |- | ||
|} <br> | |} <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'''] | ||
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.
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.
| 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
F2622 + E0240 (ligated/colonies coming up)
+ 
S03632 (ligations needed)
But BBa_S03632 is not available, so we will need to build our own version using the 3 parts below:
Parts: R0011 + C0261 + B0015
+ 
+ 
R0011 + F1610 (minis, frag needed/colonies coming up, minis thurs)
+ 
F2620 + E0240 (minis growing/ frag available)
+
