Team:Paris/July 25
From 2008.igem.org
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- | == | + | == DNA Extraction == |
- | * | + | * 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)) | ||
- | + | * Analysis of the extraction success by electrophoresis on 2µl of the parts extracted. | |
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+ | === 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) | |
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- | ===list of | + | |
+ | '''Results :''' | ||
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+ | -D100 --> D110 cl-1 ES : Absorbance ~20-50µg/ml ; DO 260/280 = 1,5 ; DO 260/230 = 0,0-0.20 | ||
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+ | -D110 cl-2 ES --> D120 : Absorbance ~0-10µg/ml | ||
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+ | === Determination of DNA concentration by electrophoresis === | ||
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+ | * 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. | ||
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+ | ==> conclusion : we don't succeed to detect DNA by spectrophometry or electrophoresis for the samples produced by Qiagen's protocol. | ||
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+ | ==> 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 | ||
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+ | == Culture == | ||
+ | |||
+ | in 10ml LB with adaptated antibiotics, all the Biobricks used before (MP100 --> MP122) | ||
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+ | * will be use to do minipreps | ||
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+ | == MiniPreps == | ||
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+ | * Use of Promega's protocol on all the clones cultivated on the 24th. | ||
+ | * Preparation of 50µl of minipreps in simple. | ||
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+ | |||
+ | |||
+ | ===list of Minipreps=== | ||
{| border="1" | {| border="1" | ||
|align="center"|'''Name''' | |align="center"|'''Name''' | ||
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|align="center"|'''Biobrick''' | |align="center"|'''Biobrick''' | ||
- | |align="center"|''' | + | |align="center"|'''Description''' |
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|align="center"|MP100 | |align="center"|MP100 | ||
- | |align="center"|B0034 | + | |align="center"|[http://partsregistry.org/Part:BBa_B0034 B0034] |
- | |align="center"| | + | |align="center"|Strongest RBS (Efficiency = 1) |
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|align="center"|MP116 | |align="center"|MP116 | ||
- | |align="center"|J23100 | + | |align="center"|[http://partsregistry.org/Part:BBa_J23100 J23100] |
- | |align="center"| | + | |align="center"|Strong constitutive promoter in J61002 |
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|align="center"|MP120 | |align="center"|MP120 | ||
- | |align="center"|B0030 | + | |align="center"|[http://partsregistry.org/Part:BBa_B0030 B0030] |
- | |align="center"| | + | |align="center"|Strong RBS (Efficiency = 0,6) |
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|align="center"|MP121 | |align="center"|MP121 | ||
- | |align="center"| | + | |align="center"|[http://partsregistry.org/Part:BBa_E0422 E0422] |
- | + | |align="center"|ECFP (RBS+LVA+Term) | |
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- | + | |align="center"| MP122 | |
- | + | |align="center"|[http://partsregistry.org/Part:BBa_E0840 E0840] | |
- | | | + | |align="center"|gfp tri-part; strong rbs |
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|} | |} |
Latest revision as of 15:32, 31 July 2008
DNA Extraction
Determination of DNA concentration by spectrophotometryConditions:
Results :
-D110 cl-2 ES --> D120 : Absorbance ~0-10µg/ml
Determination of DNA concentration by electrophoresis
Culturein 10ml LB with adaptated antibiotics, all the Biobricks used before (MP100 --> MP122)
MiniPreps
list of Minipreps
|