Biomod/2012/HKBU/BU Magician:Labook:Notebook/Labbook/2012/09/12

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2012/09/12

Overall Plan

(In the following table, “*” indicates “with”, while “X” means “without”.)
Image:HKBU2012Table1.jpg Remarks:

  • 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.


Solution Preparation

(1) Nanoparticle solution preparation
0.01g of nanoparticle solids is dissolved with 2ml HBSS (Concentration: 5mg/ml)
Further dilution:
Image:HKBU2012Table7.jpg
Total volume 2ml for each concentration.

(2) SiRNA containing solution preparation:
For each well, 300μl of nanoparticle solution is needed.
For SiRNA containing nanoparticle solution
Total volume:
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
Image:HKBU2012Table3.jpg
( *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)
=1200 μl

Take in account of pipetting error, 1200 μl-> 1600 μl
Image:HKBU2012Table4.jpg

(3)Conjugation
After adding SiRNA to both nanoparticle and HBSS solution, the tubes are stored in 4 centigrade refrigerator to allow conjugating overnight for 24hrs.

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