Team:Newcastle/10 September 2010

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=''yne''A=
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=Characterization of ''yneA''=
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Nanodrop results for pSB1C3 + ''yne''A:
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==Aim==
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#158.7
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The aim of this experiment is to prepare slides for the ''Bacillus subtilis'' 168 cells which have ''yneA'' on the plasmid (pMutin4 and pMap65) and under ''lacI'' regulation and are induced by IPTG at various concentrations.
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=Arabinose-inducible promoter characterisation=
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==Materials and Protocol==
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...
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==Miniprep of BBa_I13401 (GFP coding sequence + terminator)==
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Please refer to: [[Team:Newcastle/IPTG INduction|Slide preparation for IPTG induced cells]] for materials and protocol.
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..from last night's overnight cultures from the 8th of september transformation.
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For this experiment we used the following cell colonies:
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#''Bacillus subtilis'' 168
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#''Bacillus subtilis'' 168 with pMutin4 having ''yneA'' insert
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#''Bacillus subtilis'' 168 tith pMap65 having ''yneA'' insert
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==Double digests==
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Today we see the cells on a flourescent microscope.
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.. double digests for standard BioBrick assembly.. cut the arabinose-inducible promoter BioBrick with EcoRI and SpeI, cut gfp+terminator BioBrick with EcoRI and XbaI, for later ligation..
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==Gel extraction of digests==
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==Ligation==
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==Result and Conclusion==
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Please refer to: [[Team:Newcastle/Filamentous_Cells#Characterisation|Characterisation]] for the result.
{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 03:14, 26 October 2010

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Contents

Characterization of yneA

Aim

The aim of this experiment is to prepare slides for the Bacillus subtilis 168 cells which have yneA on the plasmid (pMutin4 and pMap65) and under lacI regulation and are induced by IPTG at various concentrations.

Materials and Protocol

Please refer to: Slide preparation for IPTG induced cells for materials and protocol. For this experiment we used the following cell colonies:

  1. Bacillus subtilis 168
  2. Bacillus subtilis 168 with pMutin4 having yneA insert
  3. Bacillus subtilis 168 tith pMap65 having yneA insert

Today we see the cells on a flourescent microscope.

Result and Conclusion

Please refer to: Characterisation for the result.

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