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?
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#2
badcell
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Posted 01 February 2005 - 03:00 AM
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 . 
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george@CASE
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Posted 02 February 2005 - 10:51 AM
badcell is right. no need to do anything with the plasmid. Sequencing is like a PCR reaction only with just one primer.
