Materials
- RNA samples
- TAE buffer
- Agarose
- Ethidium Bromide (EtBr) (10mg/mL)
- DNA loading buffer (10X)
- 1.9mM xylene cyanol, 1.5mM bromophenol blue, 25% glycerol in sterile dH2O
- DNA ladder (1kb)
Agarose Gel preparation
- Add 1.0% w/v agarose to 1x TAE buffer (e.g. 0.5g in 50mL)
- incubate at room temperature for 5 min, with occasional swirling
- Heat the suspension to melt the agarose
- Allow solution to cool before adding ethidium bromide to a final concentration of about 0.5µg/mL
- Pour solution into gel mold and allow the ‘agarose gel’ to solidify
Load and run RNA samples
- Load RNA sample mixed with 10x DNA loading buffer to a final concentration of 1x.
- Load DNA ladder
- Electrophorese gel in 1x TAE buffer at 100V for approximately 35 min.
00:35:00
- Visualize bands by UV transillumination
Determine if 28S & 18S bands are present. Use gel image analyzer to determine ratio if needed.