I am using an Invitrogen Midi prep Kit, however, I always get a very low concentration even though my initial volume is around 10-25ml.
increase your volume to 100 ml and increse solutions and incubation time accordingly and elute with less amount around 30-40micro liter.
still if u face any problem isolate it from ecoli becoz from agro its difficult and yeild and purity is also very less.
Hi Christian,
The plasmid is permitted to replicate in Agrobacterium, but with low copy number. Isolation of plasmids from Agrobacterium mostly give you low amount of DNA. Generally, people use E Coli to produce enough plasmids for digestion, sequencing and transformation. Once the digestion and sequence are verified correct; then it is used to transfer into Agrobacterium for genetic transformation. The selection marker will be used to identify those Agrobacterium colonies harboring the plasmids. There are also kits such as Qiagen Maxi and Mega prep kits available, if you really need to isolate high quantity of plasmids from Agrobacterium.
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