Sequencing of Plasmids - (Feb/01/2005 )
Hi! How do you sequence an insert in a Plasmid? Do you make a nick? How? Via Enzymes or in an ultrasoundbath?
How much ng of the plasmid do you use for the squencing reaction?
-Trinity-
You can use the supercoiled plasmid, you don't need to cut it. As few as 10-50 ng are enough, but I usually add 1 ul straight of the miniprep, without even OD'ed it .
-badcell-
badcell is right. no need to do anything with the plasmid. Sequencing is like a PCR reaction only with just one primer.
-george@CASE-