Low copy number of plasmid
Posted 27 November 2004 - 07:46 AM
I have ligated DNA target (that might be toxic for cell) to pGST-4T-2 plasmid. But, copy number of the recombinant plasmid was very low. Does anyone can help me how to get the enough amount of the plasmid after mini preparation.
Posted 27 November 2004 - 08:14 AM
1. u can try to grow cells in terrific broth
2. check the insert size limitations of the plamid
3. ensure your selction pressure conc. (antibiotic)
Posted 27 November 2004 - 09:08 AM
Thank a lot
I cultured plasmid-bearing E. coli in 4 ml of LB medium (with 4 mikroliter of 50 mg Amp). The size of insert is 200 bp.
Posted 30 November 2004 - 12:58 AM