Difference between revisions of "Team:William and Mary/Protocols"

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<b><div style = 'padding-right: 190px; padding-left: 190px;' >Protocols</div></b>
 
<b><div style = 'padding-right: 190px; padding-left: 190px;' >Protocols</div></b>
 
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<div style = 'padding-right: 190px; padding-left: 190px; line-height: 30px;' >FLow Cytometry - We used the BioRad S3e Cell Sorter to carry out single-cell fluorescence measurements. We diluted 100ul of cell culture into 400ul of Phosphate Buffered Saline (PBS). For our measurements with inductions, we split and induced the cells with IPTG 6 hours after they were inoculated. Four hours after the cells were induced, we diluted the cells in order to obtain an ideal concentration of OD 0.01. The the cells were grown for another five hours and taken out every 20 minutes to be filtered on ice and FACSed immediately. Every 100 minutes, we diluted the cells 1:10 in pre-warmed culture. We used FlowCal for downstream analysis and conversion to absolute fluorescence units.</div>
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<div style = 'padding-right: 190px; padding-left: 190px; line-height: 30px;' >FLow Cytometry - We used the BioRad S3e Cell Sorter to carry out single-cell fluorescence measurements. We diluted 100ul of cell culture into 400ul of Phosphate Buffered Saline (PBS). For our measurements with inductions, we split and induced the cells with IPTG 6 hours after they were inoculated. Four hours after the cells were induced, we diluted the cells in order to obtain an ideal concentration of OD600 0.01. The cells were then grown for another five hours and taken out every 20 minutes to be filtered on ice and FACSed immediately. Every 100 minutes, we diluted the cells 1:10 in pre-warmed culture. We used FlowCal for downstream analysis and conversion to absolute fluorescence units.</div>
  
  

Revision as of 23:50, 31 October 2017


Protocols
FLow Cytometry - We used the BioRad S3e Cell Sorter to carry out single-cell fluorescence measurements. We diluted 100ul of cell culture into 400ul of Phosphate Buffered Saline (PBS). For our measurements with inductions, we split and induced the cells with IPTG 6 hours after they were inoculated. Four hours after the cells were induced, we diluted the cells in order to obtain an ideal concentration of OD600 0.01. The cells were then grown for another five hours and taken out every 20 minutes to be filtered on ice and FACSed immediately. Every 100 minutes, we diluted the cells 1:10 in pre-warmed culture. We used FlowCal for downstream analysis and conversion to absolute fluorescence units.