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Preparing chemically competent cells: Difference between revisions
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Preparing chemically competent cells (view source)
Revision as of 20:48, 20 January 2006
, 20 January 2006→Preparation
Barry Canton (talk | contribs) |
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#Resuspend in chilled [[TSS]] buffer. The volume of [[TSS]] to use is 10% of the culture volume that you spun down. You may need to vortex gently to fully resuspend the culture, keep an eye out for small cell aggregates even after the pellet is completely off the wall. | #Resuspend in chilled [[TSS]] buffer. The volume of [[TSS]] to use is 10% of the culture volume that you spun down. You may need to vortex gently to fully resuspend the culture, keep an eye out for small cell aggregates even after the pellet is completely off the wall. | ||
#Add 100 μl aliquots to your chilled eppendorfs and store at <math>-80^o</math>C. | #Add 100 μl aliquots to your chilled eppendorfs and store at <math>-80^o</math>C. | ||
#*The original paper <cite> chung </cite> suggests freezing the cells immediately using a dry ice bath. I (BC) have used liquid nitrogen quite successfully instead of dry ice. Simply placing the cells at <math>-80^o</math>C also seems to work well ([[User:Jkm|Jkm]]) | #*The original paper <cite> chung </cite> suggests freezing the cells immediately using a dry ice bath. I ([[Barry Canton|BC]]) have used liquid nitrogen quite successfully instead of dry ice. Simply placing the cells at <math>-80^o</math>C also seems to work well ([[User:Jkm|Jkm]]) | ||
==Related topics & References== | ==Related topics & References== | ||
