sequencing

[pedro21101]

Hello
If I have DNA on wich is gene which i want to be sequenced for example by Sanger method there I need one-stranded DNA, which i need to know is how i separate that double stranded DNA to one stranded molecules (this i know) and how i distinguish that strand on which is that gene (this i dont know). Cause i separate that two strands apart but i need only one on which is gene, for sequence reaction, the other is not of my interest.
Maybe supid question, but i only begin today with it so maybe tomorow i will know it yet.
Thanks thanks thanks

[canalon]

All those steps are in the sequencing reaction. First you "melt" the double helix by boiling (or heating at 94ºCin a thermocycler). Then since you are extending a primer which is specific, if everything is OK you have your sequence.