Creating and running gels
Gel protocol for dummies.
1x TBE buffer:
- Dilute 100ml of 10x TBE buffer into 900 ml of dH20.
Reagents and instruments:
- SYBR safe gel stain
- DNA ladder
- Electrophoresis tank with plates
To create 55 ml of 1% agarose buffer:
- Add 0.55g of agarose to 55 ml of 1x TBE buffer.
- Swirl and heat for 2-4 minutes in microwave.
Note: It is a good idea to microwave for 30-45sec, stop and swirl, and then continue towards a boil.
- Let solution cool to around 50 °C and add 5 μL of SYBR Safe gel stain.
Note: Presumable more safe the ETBr, but anything that binds DNA should be treated as a carcinogen.
- Pour in plate and let rest for 1 hour.
Note: Or place in 4°C for 15-20 min.
- Mix 10 μL of sample with 5 μL of loading dye
- Transfer 15 μL to gel in electrophoresis tank and cover gel in TBE buffer.
- Run for around 60 min at 80-150V V.
Note: The DNA is negatively charged and will run towards the positive electrode. Always Run to Red.
Note: Check the run by looking at the band - stop when it's around 2/3 down.
- Visualize gel on ?