User:University of Washington/5 September 2008

From 2008.igem.org

(Difference between revisions)
(LuxR from AraC and TetR(Faifan))
 
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!0%, 0 ul!!0.05%, 1ul!!0.1%, 2ul!!0.50%, 10ul!!1.00%, 20ul
!0%, 0 ul!!0.05%, 1ul!!0.1%, 2ul!!0.50%, 10ul!!1.00%, 20ul
|-
|-
-
!rowspan=5 valign=center|aTc(ng/ml), 10ug/ml-aTc added(ul)!!0, 0ul||||||||||
+
!rowspan=5 valign=center|aTc(ng/ml), 10ug/ml-aTc added(ul)!!0, 0ul||A1||A2||A3||A4||A5
|-
|-
-
!50, 1ul||||||||||
+
!50, 1ul||B1||B2||B3||B4||B5
|-
|-
-
!100, 2ul||||||||||
+
!100, 2ul||C1||C2||C3||C4||C5
|-
|-
-
!500, 10ul||||||||||
+
!500, 10ul||D1||D2||D3||D4||D5
|-
|-
-
!1000, 20 ul||||||||||
+
!1000, 20 ul||E1||E2||E3||E4||E5
|}
|}
*added dH2O to final the volume to 200 ul
*added dH2O to final the volume to 200 ul
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*set the machine to run 24 hrs, read every 10 mins
*set the machine to run 24 hrs, read every 10 mins
 +
== Conjugation (Scott)==
 +
[[Team:University_of_Washington/Protocols#Method_.232| Conjugation method #2]]<br>
 +
·pAC99 + pUB307
 +
[[UW Glycerol Stocks| Glycerol Stock]]<br>
 +
·pAC99<br>
 +
·pAC99 + pUB307
----
----
Back to [[Team:University_of_Washington/Notebook#Notebook]]
Back to [[Team:University_of_Washington/Notebook#Notebook]]

Latest revision as of 19:36, 10 September 2008

LuxR from AraC and TetR(Faifan)

- measured Absorbance of the overnight culture at 660 nm = 1.5

- diluted 60 ul culture into 5.40 ml Tsy+KAN. Absorbance at 660 nm = 0.035

- incubated 37 degree Celsius for 2.5 hrs, Absorbance at 600 nm = 0.7

- diluted 1 ml culture into 5 ml Tsy+Kan, OD@600nm = 0.165

- set up 96 well plate reader experiment:

  • 5x5, 160 ul culture
Arabinose(%), 10%Arabinose added (ul)
0%, 0 ul0.05%, 1ul0.1%, 2ul0.50%, 10ul1.00%, 20ul
aTc(ng/ml), 10ug/ml-aTc added(ul)0, 0ulA1A2A3A4A5
50, 1ulB1B2B3B4B5
100, 2ulC1C2C3C4C5
500, 10ulD1D2D3D4D5
1000, 20 ulE1E2E3E4E5
  • added dH2O to final the volume to 200 ul
  • added 40 ul of mineral oil to each well to protect evaporation
  • set the machine to run 24 hrs, read every 10 mins

Conjugation (Scott)

Conjugation method #2
·pAC99 + pUB307

Glycerol Stock
·pAC99
·pAC99 + pUB307


Back to Team:University_of_Washington/Notebook#Notebook