Even if I get them cultured colonies are very less and almost not transferable and grows very slowly in the media.
I checked cells by transforming with control vector.
I changed antibiotic also which I am using to screen my colony.
I checked the incubator for the temperature.
I checked ligation using control ligation.
Also I checked both the digestion separately.
Everything gave positive results.
Can anybody suggest any possible solution to this.
If your control ligation is fine, then the protocol should be fine. Try to check the molar ratio of vector and inserts. If you have colleagues doing transformation with 1) the same vector as the one you are using, 2) have inserts of similar size as yours and 3) using the same protocol as yours; if they get successful colonies with the desired inserts, then the problem lies with your vector-insert construct.
What kind of gene did you transferred? if the protein produced is toxic, maybe the cells were killed. I had some experience with it. If so, you may consider adding glucose to inhibit expression or reduce the temperature. Good luck.
As suggested by Heru Pramono, it seems likely that your construct expresses a toxic polypeptide. You may avoid the problem by choosing a strain that affords stringent control of expression, such as BL21AI, which is inducible by L-arabinose. It provides a more stringent repression than BL21 (DE3), even with the pLysS plasmid.
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