Team:Rice University/Notebook/12 June 2008

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=Thursday 12 June=
=Thursday 12 June=
*Taylor Stevenson
*Taylor Stevenson
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**Transformation From New Registry (Paper Punches)-[[Team:Rice University/Electroporation|Electroporation]] and [[Team:Rice_University/Heat_Shock|Heat Shock]] of [http://partsregistry.org/Part:BBa_I744204 BBA_I744204] located on [http://partsregistry.org/Part:pSB1A2 pSB1A2] and [http://partsregistry.org/Part:BBa_I13521 BBA_I13521] located on [http://partsregistry.org/Part:pSB1A2 pSB1A2] into XL1-Blue MR electrocompetent cells.
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**Transformation From New Registry (Paper Punches)-[[Team:Rice University/Electroporation|Electroporation]] and [[Team:Rice_University/Heat_Shock|Heat Shock]] of [http://partsregistry.org/Part:BBa_I744204 BBA_I744204] located on [http://partsregistry.org/Part:pSB1A2 pSB1A2] (positive control) and [http://partsregistry.org/Part:BBa_I13521 BBA_I13521] located on [http://partsregistry.org/Part:pSB1A2 pSB1A2] into XL1-Blue MR electrocompetent and XL1-Blue chemically competent cells.
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*#Added 2uL of DNA solution obtained by soaking DNA punch in (10:1) Tris:EDTA buffer incubated @ 42*C for 20min.
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*#Added 1uL of stock miniprepped [http://partsregistry.org/Part:BBa_I744204 BBA_I744204] DNA solution and 2uL obtained by soaking DNA punch in (10:1) Tris:EDTA buffer incubated @ 50*C for 30min to both chemical and electrocompetent cells.
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*#Electropulse parameters: 1.8 kV & 5.4 ms
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*#Electropulse parameters for the miniprepped plasmid and extracted plasmid where 1.8kV & 5.2ms and 1.8kV & 5.0ms respectively.
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*#Plated 100uL and 20uL of transformed cells as well as 20uL of  1/100 and 1/10,000 dilutions in sterile 50% glycerol.  Plates were cultured O/N.  
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*#Plated 100uL of each the transformed and cultured cells on LB/Amp plates.  Plates were cultured O/N @ 37*C.
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**'''Result'''-no colonies were observed at any dilution
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**'''Result'''-no colonies were observed on plates from the extracted DNA while >>1,000 colonies grew on plates resulting from miniprepped DNA.

Revision as of 17:30, 25 June 2008

Thursday 12 June

  • Taylor Stevenson
    • Transformation From New Registry (Paper Punches)-Electroporation and Heat Shock of [http://partsregistry.org/Part:BBa_I744204 BBA_I744204] located on [http://partsregistry.org/Part:pSB1A2 pSB1A2] (positive control) and [http://partsregistry.org/Part:BBa_I13521 BBA_I13521] located on [http://partsregistry.org/Part:pSB1A2 pSB1A2] into XL1-Blue MR electrocompetent and XL1-Blue chemically competent cells.
    1. Added 1uL of stock miniprepped [http://partsregistry.org/Part:BBa_I744204 BBA_I744204] DNA solution and 2uL obtained by soaking DNA punch in (10:1) Tris:EDTA buffer incubated @ 50*C for 30min to both chemical and electrocompetent cells.
    2. Electropulse parameters for the miniprepped plasmid and extracted plasmid where 1.8kV & 5.2ms and 1.8kV & 5.0ms respectively.
    3. Plated 100uL of each the transformed and cultured cells on LB/Amp plates. Plates were cultured O/N @ 37*C.
    • Result-no colonies were observed on plates from the extracted DNA while >>1,000 colonies grew on plates resulting from miniprepped DNA.






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