Team:Paris/July 25

From 2008.igem.org

(Difference between revisions)
(→Determination of DNA concentration by spectrophotometry)
(→Determination of DNA concentration by spectrophotometry)
Line 22: Line 22:
D100 --> D110 cl-1 ES : Absorbance ~20-50µg/ml ;  DO 260/280 = 1,5 ;  DO 260/230 = 0,0-0.20
D100 --> D110 cl-1 ES : Absorbance ~20-50µg/ml ;  DO 260/280 = 1,5 ;  DO 260/230 = 0,0-0.20
D110 cl-2 ES --> D120 : Absorbance ~0-10µg/ml
D110 cl-2 ES --> D120 : Absorbance ~0-10µg/ml
 +
 +
 +
 +
=== Determination of DNA concentration by electrophoresis ===
* to check for the samples from Qiagen's protocol, if the absence of detection of absorbance, is due to a problem of reading or a problem during the extraction, we realise an electrophoresis.
* to check for the samples from Qiagen's protocol, if the absence of detection of absorbance, is due to a problem of reading or a problem during the extraction, we realise an electrophoresis.
Line 30: Line 34:
==> we decide to repeat all the samples from the beginning, so we culture O/N at 37°c all the strains usefull for the experiments
==> we decide to repeat all the samples from the beginning, so we culture O/N at 37°c all the strains usefull for the experiments
 +
 +
 +
== Culture ==
== MiniPreps ==
== MiniPreps ==

Revision as of 12:18, 28 July 2008

← Yesterday

↓ Calendar ↑

Tomorrow →


Contents

Extraction of the DNA

  • Use of Promega's protocol for the extraction of: gel n°1 --> 4 : (D100 --> D110 cl-1 ES))
  • Use of Qiagen's protocol for the extraction of: gel n°5 --> 8 : (D110 cl-2 ES --> D120))
  • Test of the succeed of the extraction by electrophoresis on 2µl of the parts extracted.


Determination of DNA concentration by spectrophotometry

conditions: 2µl of the DNA extracted 98µl of pure water

  • Reading against the adaptated blank (2µl of the elution's buffer / 98µl of pure water)
  • List of the results :

D100 --> D110 cl-1 ES : Absorbance ~20-50µg/ml ; DO 260/280 = 1,5 ; DO 260/230 = 0,0-0.20 D110 cl-2 ES --> D120 : Absorbance ~0-10µg/ml


Determination of DNA concentration by electrophoresis

  • to check for the samples from Qiagen's protocol, if the absence of detection of absorbance, is due to a problem of reading or a problem during the extraction, we realise an electrophoresis.


==> conclusion : we don't succeed to detect DNA by spectrophometry or electrophoresis for the samples produced by Qiagen's protocol.


==> we decide to repeat all the samples from the beginning, so we culture O/N at 37°c all the strains usefull for the experiments


Culture

MiniPreps

  • Use of Promega's protocol on all the clones cultivated on the 24th.
  • Preparation of 50µl of minipreps in simple.


list of Minipreps

Name Biobrick Description
MP100 [http://partsregistry.org/Part:BBa_B0034 B0034] Strongest RBS (Efficiency = 1)
MP116 [http://partsregistry.org/Part:BBa_J23100 J23100] Strong constitutive promoter in J61002
MP120 [http://partsregistry.org/Part:BBa_B0030 B0030] Strong RBS (Efficiency = 0,6)
MP121 [http://partsregistry.org/Part:BBa_E0422 E0422] ECFP (RBS+LVA+Term)
MP122 [http://partsregistry.org/Part:BBa_E0840 E0840] gfp tri-part; strong rbs
Retrieved from "http://2008.igem.org/Team:Paris/July_25"