u/Maleficent-Lab-1113

I am loosing too much DNA amount after gel extraction. Can anyone suggest how can I increase the yield of DNA after gel extraction? I am using Qiagen kit

Hi everyone, I had a PCR-amplified product of a 7.7 kb plasmid (Whole plasmid amplification). But after DpnI digestion, my amplified product, along with the template control, was fully digested. Why is it happening? Is there something with the DpnI enzyme itself, or was my product not amplified? I used 200 ng of the template plasmid for amplification.

I am performing a cloning where I need to replace a 2.3Kb insert from the vector (5.4Kb) with another similar-sized insert(2.37Kb). After ligation, although I am getting colonies in the ligation plates, even my vector-only (double-digested) plate shows plenty of colonies. I picked up 2-3 colonies randomly from the control plate for screening, and I got an insert release. Is it because some vector got uncut? If so how to avoid this and get the desired clone?