Editing MRNA Transfection (section)

== Generation Of Carbonate Apatite Particles<br>  ==

Addition of only 3 mM Ca2+ to the HCO3&nbsp; buffered cell culture medium (DMEM or Willium E, pH 7.5) and incubation at 370C for 30 min, resulted in microscopically visible particles. Generation of these particles only in HCO3, but not in Hepes-buffered media or solution (pH 7.5) containing the same amount of exogenous Ca2+ and phosphate (0.9 mM), indicates the possible involvement of carbonate along with phosphate and Ca2+ in particle formation.<br> 

<br> 

1. Mix 3 to 6 μl of 1 M CaCl2 with 2 μg of plasmid DNA in 1 ml of fresh serum-free HCO3- –buffered (pH 7.5) medium (DMEM)<br> 

2. Incubation for 30 min at 37 °C for complete generation of DNA/carbonate apatite particles. <br> 

3. Add&nbsp; particles to the rinsed cultured cells. 

4. Change medium after 4-h incubation 

5. Assay after 1 day.<br><br>