5.6 RT-PCR analysis

Make enough for 2 gels/pair 1.5% agarose

gels mL TAE g agarose µL EtBr
11 550 8.25 5.5
20 1000 15 10
22 1100 16.5 11
24 1200 18 12

Use EtBr TAE

Use LMW mass ruler

  • Prepare 15 μL of each of your PCR reactions for loading on gels by transferring to a clean, labeled 1.5 mL microfuge tube, and adding 3 μL of loading dye. Load the entire 18 μL – it is important that all lanes contain the same amount.

  • Load gels, and run at 100V for 45 minutes. Remember to load 5 μL (= 220 ng) of LMW Mass Ladder as your standard.

  • Analyze gels