sequencing

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pedro21101
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sequencing

Post by pedro21101 » Tue Dec 06, 2005 6:49 pm

Hello :D
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

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canalon
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Post by canalon » Tue Dec 06, 2005 8:53 pm

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.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

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