Problem:Standard curve achieved but poor discrimination between points.
Possible causes 1:Improper calculation of standard curve dilutions
Solutions 1:Check the calculation results and make new standard curve
Possible causes 2:Insufficient washing
Solutions 2:Ensure sufficient washing
Possible causes 3:Plate sealer reused
Solutions 3:Use a fresh plate sealer for each step
Possible causes 4:Not used plate sealers
Solutions 4:Use plate sealers
Possible causes 5:Buffer contaminated
Solutions 5:Prepare fresh buffer
Possible causes 6:Insufficient washing
Solutions 6:Ensure sufficient washing. If using an automatic plate washer, check that all ports are clean and unobstructed.
Possible causes 7:Variations in incubation temperature
Solutions 7:Adhere to recommended incubation temperature; Avoid putting plates in variable environmental conditions.