Brown:Transformation Protocol with Fischer cells
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4. INCUBATE ON ICE FOR 30 MIN. | 4. INCUBATE ON ICE FOR 30 MIN. | ||
| - | 5. HEAT SHOCK AT 42˚C FOR 30 SEC. | + | 5. HEAT SHOCK AT 42˚C IN WATER BATH FOR 30 SEC. |
6. PLACE ON ICE FOR 2 MIN. | 6. PLACE ON ICE FOR 2 MIN. | ||
Latest revision as of 19:27, 6 July 2006
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TRANSFORMATION PROTOCOL WITH FISCHER COMPETENT CELLS
1. TAKE CELLS FROM -80 AND THAW FOR IMMEDIATE USE
2. MIX CELLS GENTLY WITH TAPPING
3. ADD 4µL DNA TO 50µL COMPETENT CELLS (EACH VIAL CONTAINS 50µL)
4. INCUBATE ON ICE FOR 30 MIN.
5. HEAT SHOCK AT 42˚C IN WATER BATH FOR 30 SEC.
6. PLACE ON ICE FOR 2 MIN.
7. ADD 250µL CIRCLE GROW
8. INCUBATE FOR 1 HOUR WITH SHAKING AT 37˚C
9. ADD 250µL CIRLCE GROW
10. PLATE 200µL OF MIX
