Vector Vs Host - PGL2 Vs DH5alpha (Jan/16/2005 )
Hi guys,
really love this forum.
well i have PGL2 luciferase vector from promega supplied with JM109 cells in glycerol stock.
sooner i will be doing mammalian transfection studies, but QIAGEN plasmid miniprep says JM109 strain in not so good for plasmid prep and other downstream process as JM series produce large amt of carbohydrates and may inihibit enzyme activites.
now tht i also i have a stock of DH5alpha, can i use PGL2(promega) with DH5a(invitrogen). will it make a diffrence on plasmid yeild and quality?
please explain.
and how serious is the contamination of endotoxins in palsmid DNA affect mammalian transfection studies.
thx in advance.
rajgene
He Rajgene,
I have used pGL vectors and JM109 cells a lot. All my plasmid preps were done with Qiagene kit without any problem in downstream applications such as restriction digestion, transfection, subcloning. You should not worry about JM109.