Brown:Transformation Protocol with Fischer cells

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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