Calcium Phosphate Method Of Transfecting Drosophila S2 Cells Solutions:

Calcium phosphate method of transfecting Drosophila S2 cells Solutions:

- 2x HBS HEPES 10g/l NaCl 16g/l pH 7.10 with 2N NaOH, filter sterilize. pH is critical for forming good ppt. - 100x PO4: Mix equal parts of 70mM Na2HPO4 (993.7 mg/100ml) and 70mM NaH2PO4 (965.9 mg/100 ml). Filter sterilize. - 2M CaCl2. Filter sterilize. Solutions can be stored at RoomT. HEPES is good for only 1-2 months. Aliquots of HEPES can be stored in -80dC.

Preparation of precipitate: For each construct, set up two falcon tubes (12 x 75 mm) labelled A and B. Add: A. 0.5 ml of 2x HBS + 10 ul of 100x PO4 B. 440 ul H2O + 20ug DNA + 60 ul CaCl2 (add last). Flick mix . Introduce solution B dropwise into Soln A while bubbling A. Let soln. sit for 5-30 mins. Add to cells while gently mixing.