Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/02/10

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(02/10/2014)
Current revision (19:14, 10 February 2014) (view source)
(02/10/2014)
 
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|}
 +
Golden gate to build LuxR receiver
 +
 +
{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Part'''
 +
| align="center" style="background:#f0f0f0;"|'''Length (bp)'''
 +
| align="center" style="background:#f0f0f0;"|'''Concentration (ng/ul)'''
 +
| align="center" style="background:#f0f0f0;"|'''fmol/ul'''
 +
| align="center" style="background:#f0f0f0;"|'''Final volume to dilute 1ul to get 20 fmol/ul'''
 +
|-
 +
| Receiver vector||2372||113||72.2||3.6
 +
|-
 +
| Lux box||56||20||541.1||27.1
 +
|-
 +
| -35-B0033-GFP-TT||945||124||198.8||9.9
 +
|-
 +
| R0011-RBS||85||5||89.1||4.5
 +
|-
 +
| LuxR||817||57||105.7||5.3
 +
|}
 +
 +
GG to build RhlR receiver
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Part'''
 +
| align="center" style="background:#f0f0f0;"|'''Length (bp)'''
 +
| align="center" style="background:#f0f0f0;"|'''Concentration (ng/ul)'''
 +
| align="center" style="background:#f0f0f0;"|'''fmol/ul'''
 +
| align="center" style="background:#f0f0f0;"|''''''
 +
|-
 +
| Receiver vector||2372||113||72.2||3.6
 +
|-
 +
| Rhl box||56||20||541.1||27.1
 +
|-
 +
| -35-B0033-GFP-TT||945||124||198.8||9.9
 +
|-
 +
| R0011-RBS||85||5||89.1||4.5
 +
|-
 +
| RhlR||765||50||99.0||5.0
 +
|}
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Current revision

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02/10/2014

Ran gel of PCR products from 2/8/14.

Sample Primer 1 Primer 2 Template Ideal AT Approx Length
1P098P099I13522 (2)462372
2P098P099I13522 (3)462372
3P098P099I13522 9.27 55.7462372
4P103P104I13522 (2)49780
5P103P104I13522 (3)49780
6P103P104I13522 9.27 55.749780


      • insert gel photo before and after***


Gel extract using Zymo kit. Read concentrations of these and gel extractions from 2/7/14:

Primer 1 Primer 2 Template 260 280 260/280 ng/µL
water0.0030.0021.9333.084
P103P104I13522 (2)0.1240.0651.9124.369
P103P104I13522 (3)0.1230.0651.9123.324
P103P104I13522 9/27/13 55.70.0540.0281.91854.117
P098P099I13522 (3)0.0980.0511.93197.574
P098P099I13522 (2)0.1160.061.943115.692
P117P118K092900 (RhlR)0.050.0271.87450.159
P105P106R0011 (2)0.0050.0022.6325.229
P113P114LuxR (F2620)0.0570.0291.96857.356
P105P106R0011 (1)0.0020-6.6672.073
P088P102Receiver vector0.1130.061.896113.028

Golden gate to build LuxR receiver

Part Length (bp) Concentration (ng/ul) fmol/ul Final volume to dilute 1ul to get 20 fmol/ul
Receiver vector237211372.23.6
Lux box5620541.127.1
-35-B0033-GFP-TT945124198.89.9
R0011-RBS85589.14.5
LuxR81757105.75.3

GG to build RhlR receiver

Part Length (bp) Concentration (ng/ul) fmol/ul '
Receiver vector237211372.23.6
Rhl box5620541.127.1
-35-B0033-GFP-TT945124198.89.9
R0011-RBS85589.14.5
RhlR7655099.05.0


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