Thank you for valuable suggestion.

Hello Pravin,

You can also check your medium before you throw it by keeping it in to incubator 5%CO2+37 degree C for 24 hour. After that if media is contaminated you can see contamination in media bottle. Also, good way to take everything new and try to be careful in each step of cell culture work. May be its good idea to take everything new and sterilized everything once more to be confirm that every thing is sterilized and then you can start new passage of cell..Good luck!