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Optimizing electroporation parameters

Irrespective of the buffer system selected, it is essential to ensure that the cells do not remain in the electroporation buffer for longer than 30 minutes.

It is recommended to transfer the cells 510 minutes after the pulse into growth medium and incubate them for at least 23 hours at 37C before any centrifugation takes place. This step markedly increases the survival rate.

Desired osmolarity Eppendorf Hypoosmolar Buffer (ml) Eppendorf Isoosmolar Buffer (ml)
90 mOsmol/kg 10.0 0.0
150 mOsmol/kg 6.8 3.2
200 mOsmol/kg 4.2 5.8
250 mOsmol/kg 1.6 8.4
280 mOsmol/kg 0.0 10.0
Table 2.<
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