Green Lab Nanoparticle For 6 Well Cardiomyocyte Transfection
Edwin Yoo
Published: 2023-02-23 DOI: 10.17504/protocols.io.ewov1ojd2lr2/v1
Abstract
nanoparticle transfection
Steps
Nanoparticle Synthesis
1.
DNA Dilution
For 3 replicate wells of one condition in 6-well plate format, mix the following in microcentrifuge tube.
- 60 ul DNA at 1ug/ul
- 440 ul sodium acetate
- Total volume = 500 ul for 3 replicate wells per one condition
2.
Polymer Dilution (7.2 mg.ml polymer concentration in NaAc)
For 3 replicate wells of one condition in 6-well plate format, mix the following in microcentrifuge tube.
- 40 ul polymer (e.g. 4-5-6)
- 460 ul sodium acetate
- Total volume = 500 ul for 3 replicate wells per one condition
3.
Nanoparticle Synthesis
Combine diluted DNA with diluted polymer
- Add 500 ul of diluted DNA to 500 ul diluted polymer (can freeze here at -80 at your own risk)
- Add 5000 ul of media to 1000 ul NP's for final volume master mix of 6000 ul per condition
- Incubate for 10 minutes
4.
Transfection
- Change media from each well of 6 well plate with 2000 ul nanoparticle master mix (media + nanoparticles)
- Incubate for 24 hours or desired time
