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Team:Rice University/Notebook/12 June 2008
From 2008.igem.org
Thursday 12 June
- Taylor Stevenson
- Transformation From New Registry (Paper Punches)-Electroporation and Heat Shock of BBa_I744204 located on pSB1A2 (positive control) and BBa_I13521 located on pSB1A2 into XL1-Blue MR electrocompetent and XL1-Blue chemically competent cells.
- Added 1uL of stock miniprepped BBa_I744204 DNA solution and 2uL obtained by soaking BBa_I13521 DNA punch in (10:1) Tris:EDTA buffer incubated @ 50*C for 30min to both chemical and electrocompetent cells.
- Electropulse parameters for the miniprepped plasmid and extracted plasmid where 1.8kV & 5.2ms and 1.8kV & 5.0ms respectively.
- 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.
- Obtaining a lambda lysogen
- VCS257 cells were prepared for phage infection as specified in packaging manual [1] and infected with phage at roughly a 1/10 pfu/cfu ratio. Resulting phage/bacteria mixture was incubated @ 37*C for 20 min and then streaked onto an LB plate. Plate was incubated @ 30*C O/N.
- Result-approximately 100 possibly lysogenic colonies grew on plate.
- Transformation From New Registry (Paper Punches)-Electroporation and Heat Shock of BBa_I744204 located on pSB1A2 (positive control) and BBa_I13521 located on pSB1A2 into XL1-Blue MR electrocompetent and XL1-Blue chemically competent cells.
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