other misc resources

> Science Journals

> Cell Culture Handbook

> Cell contamination

> fluorochrome table

> stable transfection

other notes:

1000-1500 rpm for 5 mins....for freezing cells

Tissue culture media contains Calcium and Magnesium ions, foetal calf serum contains proteins that are trypsin inhibitors. Both Mg2+/Ca2+ INHIBIT TRYPSIN. The reason why we use PBS without Ca2+/Mg2+ to wash the cells prior to trypsinisation is to reduce the concentration of Divalent cations and proteins that inhibit trypsin action. EDTA is a Calcium chelator which will "mop" up the remaining divalent cations. If trypsin is allowed to stay in contact with the cells for too long a time, cell viabilty will reduce.

This should be the first principle of cell culture that you learn on day one. There are only very few cells that will detach with EDTA treatment alone.


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