High CV from Bubbles in Wells (ELISA (Inconsistent Results / High CV))

Question

Accepted Answer

Inconsistent absorbance readings between replicate wells with coefficient of variation >15%. Visual inspection may reveal bubbles present in wells, particularly when using detergent-containing solutions. Common causes: 1) Solutions containing detergents are especially susceptible to bubble formation 2) Rapid or forceful addition of reagents to wells introduces air 3) Bubbles remaining in wells interfere with optical reading path 4) Multi-channel pipettors may trap air when dispensing detergent solutions Solutions: 1) Ensure no bubbles are present prior to reading the plate by visual inspection 2) Gently add solutions to the wells to minimize turbulence and air incorporation 3) Pop bubbles by touching them with a dry pipette tip before reading 4) Allow plate to stand 1-2 minutes after reagent addition for bubbles to dissipate