Hi guys, I am very confused at the moment. I would like to check the membrane integrity of bacteria after exposure of antibiotic. I filtered my samples after incubation and check the absorbance at 260. All my samples even plain PBS or water had value of 3.2, which is very weird. I use clear 96 wellplate for my measurement. Just wondering do you guys encounter this problem before? Why does miliq water/PBS have value at A260? Thank you.
Are you sure your plates are truly UV transparent at 260? Regular microplates do not let UV through and absorb very heavily, even many UV transparent plates are only transparent down to 300 or 320 but not 260. I would check your PBS in a regular spectrophotometer with UV cells and compare to the reading for your plates but I think that is your problem.
Are you sure that the 96 wellplate you use is transparent to the 260nm ultraviolet spectrophotometer?
It is recommended that you test the blank board once!
Unless you are using optical plates, 260nm will likely provide distorted results. Have you tried reading empty/medium only wells?
- Badges
- Science method