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

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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
1 P098 P099 I13522 (2) 46 2372
2 P098 P099 I13522 (3) 46 2372
3 P098 P099 I13522 9.27 55.7 46 2372
4 P103 P104 I13522 (2) 49 780
5 P103 P104 I13522 (3) 49 780
6 P103 P104 I13522 9.27 55.7 49 780


      • 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
water 0.003 0.002 1.933 3.084
P103 P104 I13522 (2) 0.124 0.065 1.9 124.369
P103 P104 I13522 (3) 0.123 0.065 1.9 123.324
P103 P104 I13522 9/27/13 55.7 0.054 0.028 1.918 54.117
P098 P099 I13522 (3) 0.098 0.051 1.931 97.574
P098 P099 I13522 (2) 0.116 0.06 1.943 115.692
P117 P118 K092900 (RhlR) 0.05 0.027 1.874 50.159
P105 P106 R0011 (2) 0.005 0.002 2.632 5.229
P113 P114 LuxR (F2620) 0.057 0.029 1.968 57.356
P105 P106 R0011 (1) 0.002 0 -6.667 2.073
P088 P102 Receiver vector 0.113 0.06 1.896 113.028

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 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

Part Length (bp) Concentration (ng/ul) fmol/ul '
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