OhioMod2013:Results: Difference between revisions
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===HPLC=== | ===HPLC=== | ||
Show A280 peaks, determine rate of DNA encapsulation vs free DNA. | Show A280 peaks, determine rate of DNA encapsulation vs free DNA. | ||
===Ca:P ratio=== | |||
Use inductively coupled mass spectroscopy to determine Ca:P ratio. | |||
===Zetasizer=== | ===Zetasizer=== | ||
Determine size and zeta potential of particles. | Determine size and zeta potential of particles. | ||
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==Serum Assay== | ==Serum Assay== | ||
Apply DNA encapsulated particles vs free DNA origami vs PEGylated particles to bovine serum. | Apply DNA encapsulated particles vs free DNA origami vs PEGylated particles to bovine serum. | ||
Measure intact origami after period of time. | Measure intact origami after period of time via agarose gel electrophoresis. | ||
Apply endonucleases/DNase to encapuslated vs free DNA origami. | |||
Measure intact orgimai after periods of time via agarose gel elctrophoresis. | |||
==Cytotoxicity== | ==Cytotoxicity== | ||
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Use chloroquinine to inhibit lysosomal fusion/acidification. Measure effect. | Use chloroquinine to inhibit lysosomal fusion/acidification. Measure effect. | ||
May also use Cytochalasin D to inhibit endosome travel. Measure effect | |||
Measure transfection of confluent cells vs recently split fast-growing cultures. | |||
Revision as of 10:57, 4 May 2013
Results
Nanoparticle Characterizatione
HPLC
Show A280 peaks, determine rate of DNA encapsulation vs free DNA.
Ca:P ratio
Use inductively coupled mass spectroscopy to determine Ca:P ratio.
Zetasizer
Determine size and zeta potential of particles.
TEM
Take direct measurements of ~100 particles. Determine consistency of size.
Dissolution
Perform titration of nanoparticles until dissolution of nanoparticles. Run gel for DNA origami vs. free DNA after dissolution. Is origami intact?
Serum Assay
Apply DNA encapsulated particles vs free DNA origami vs PEGylated particles to bovine serum. Measure intact origami after period of time via agarose gel electrophoresis.
Apply endonucleases/DNase to encapuslated vs free DNA origami. Measure intact orgimai after periods of time via agarose gel elctrophoresis.
Cytotoxicity
Perform MTT assay.
Cellular Uptake
Use carbazole-based biscyanine fluorophores to measure intact DNA origami intracellularly. Also used flourescent tagged DNA origami. Also use flourescent tagged Calcium Phospahte nanoparticles. potential for confocal microscopy.
Use chloroquinine to inhibit lysosomal fusion/acidification. Measure effect. May also use Cytochalasin D to inhibit endosome travel. Measure effect
Measure transfection of confluent cells vs recently split fast-growing cultures.
