IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/07/30

From OpenWetWare
Jump to: navigation, search
Igem-logo-150px.png Memo-Cell Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png


  • Insert your content here.
Igem-logo-150px.png Population counter Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Aleksandra
Gel purification with the Quiagen kit
Purification of the bands extracted yesterday :

  • term/attC/mRFP
  • weak.RBS /TetA(C)
  • med.RBS /TetA(C)
  • stand.RBS /TetA(C)
  • weak.RBS /LacZ
  • stand.RBS /LacZ


Léa
PCR purification with the Quiagen kit

  • attC
  • LuxI/term


Léa, Aleksandra and Raphaël
Nanodrop
Quantification of DNA amounts in the minipreps from yesterday

Sample name 260/280 260/230 ng/µl
stand.RBS /TetA(C) 1 1.93 2.31 271.3
pLux/RBS/GFP/term A 1.90 2.18 244.8
Pmed/weakRBS/LuxR 1 1.91 1.52 87.8
Pstr/weakRBS/LuxR 1 1.94 2.28 88.4
Pmed /medRBS/LuxR 1 1.89 2.19 156.2
Pstr/medRBS/LuxR 1 1.91 2.04 83.9
Pmed/strRBS/LuxR 1 1.94 2.37 236.5
Pstr/strRBS/LuxR 1 1.96 2.43 101.3
stand.RBS/LacZ 1 1.88 1.68 224.7
weak RBS/LacZ 1 1.92 2.06 162.4
LuxI/term 1 1.91 2.18 325.2
attC 1.97 1.96 152.0
term/attC/mRFP 2.03 2.36 213.2


Quantification of DNA amounts in the gel purification and PCR purification samples

Sample name 260/280 260/230 ng/µl
term/attC/mRFP (gel purif.) 1.34 0.47 9.3
weak.RBS /TetA(C) (gel purif.) 1.67 0.64 27.0
med.RBS /TetA(C) (gel purif.) 1.90 0.54 24.2
stand.RBS /TetA(C) (gel purif.) 1.76 0.52 31.3
weak.RBS /LacZ (gel purif.) 1.45 0.47 11.8
stand.RBS /LacZ (gel purif.) 1.85 0.07 4.0
attC (PCR purif.) 1.42 0.70 19.1
LuxI/term (PCR purif.) 1.60 2.03 22.1
attC (PCR purif. from 07/23) 1.69 1.24 22.4


Aleksandra
Ligation using around 150ng of the insert and 50µg of the vector.
Incubation 1h at 25°C.

  • 2µl attC (07/23) + 15µl term/attC/mRFP + 2µl buffer + 0.5µl ligase
  • 2µl LuxI/term + 6µl weak.RBS /TetA(C) + 1µl buffer + 0.5µl ligase
  • 2µl LuxI/term + 6µl med.RBS /TetA(C) + 1µl buffer + 0.5µl ligase
  • 2µl LuxI/term + 6µl stand.RBS /TetA(C) + 1µl buffer + 0.5µl ligase
  • 2µl LuxI/term + 15µl weak.RBS /LacZ + 2µl buffer + 0.5µl ligase
  • 2µl LuxI/term + 30µl stand.RBS /LacZ + 3.6µl buffer + 0.5µl ligase

Raphaël
Transformation in TOP10 cells

  • attC/term/attC/mRFP
  • weak.RBS /TetA(C)/LuxI/term
  • med.RBS /TetA(C)/LuxI/term
  • stand.RBS /TetA(C)/LuxI/term
  • weak.RBS /LacZ/LuxI/term
  • stand.RBS /LacZ/LuxI/term
  • pUC 19 positive control


→ there are colonies on every plate