- 3,6 have not been conducted due to the limited number of magnets.
- 7 has not been conducted due to the same reason.
- Cell with 1mg/ml nanoparticle without magnetic field has been conducted in the following experiments.
(2) SiRNA containing solution preparation:
For each well, 300μl of nanoparticle solution is needed.
For SiRNA containing nanoparticle solution
300x 2(duplicates) x2(with magnetic field/without magnetic field)=1200μl
Take in account the pipetting error, 1200μl -> 1600μl for each concentration solution containing nanoparticles
SiRNA stock concentration:40μM
SiRNA concentration needed for treatment 200nM
( *Remarks:The final concentration of SiRNA containing nanoparticle solution is not calculated here.)
For SiRNA containing HBSS solution (experiment 7,8)
Volume of SiRNA containing HBSS
= 300 μlx2 (duplicates) x 2(experiment7,8)
Take in account of pipetting error, 1200 μl-> 1600 μl
After adding SiRNA to both nanoparticle and HBSS solution, the tubes are stored in 4 centigrade refrigerator to allow conjugating overnight for 24hrs.