thank you for your opinion, the purpose of co-transformation is to insert another plasmid into a host which is will help my vector-insert to be expressed well. Right now, i already got it by performing standard transformation with combination of two plasmid in less than 10% of competent cell volume..thank you very much
do you means i still need redo the transformation,or just optimize the extraction for restriction enzymes digestion?some of my colleague said these happen because of insufficient concentration of DNA that need for the digestion..
Is it a high copy number vector ?
Your ligation sounds fine to me
Is it because your insert DNA too toxic for the cell because of the high copy number vector?
It's just a thought
what you mean by toxic?
i really need a clearance, i'm still a newbie in this area
my insert size is 1.5k bp..