I inoculated transformed E.coli on these plates, after electroporation for blue white selection.
1-Medium temperature was higher during spills in plates
2-Agar concentration was low and the water is collected on the plate.
3-May be Incubation temperature was high
4-Source of used agar is not good.
There are so many reasons like -
There might be some moisture absorbances.
If you will desiccator or you will perform your experiment your experiment in dry condition.
Also, try to make surrounding moisture free.
Use dried plates only.
Most importantly the RATIO of agar and liquid used should be according to the protocol, because the extra unused moisture may cause the situation what you asked for.
Dear Himanshi,
first: check your recipe (proportion of agar/water; expiring date of your agar, organoleptic properties, etc) and check the temperature control of your incubator.
then, be sure to have properly casted your plates, if they are kept cold, let them get RT; if they are hot, allow them to dry properly before starting the experiment.
Then, are you sure you have been careful and kind when you inoculated your bugs, letting them to spread and penetrate the agar, toghether with the liquid?
Then, have you incubated the plates with parafilm seal (you should not, parafilm is for avoiding dissecation while storing)? Have you incubated them upside down?
I am sorry if I am being exceedengly motherly, I have commited all of these wrongdoings myself when starting cloning!
Good luck!
Rocio
@Rocio Hassan..no probs, the chemicals are ok i.e. agar and all, but I doubt Ampicillin, next, yes I sealed my plates with parafilm. So, maybe next time I won't seal the plates what say? and yes I incubated the plates upside down.
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