BC basic - from big bang

1
How DNA Sequence Is Determined?
DNA fragments having a difference of one
nucleotide can be separated on gel
electrophoresis
Polyacrylamide Gel Electrophoresis
T A G C
G
C
T
A
But these bands cant tell us the identity of the
terminal nucleotides
If those band with the same terminal nucleotide
can be grouped, then it is possible to read the
whole sequence
Juang RH (2004) BCbasics
2
How to Obtain DNA Fragments
Chemical method
AT
ATCGAT
Non-radioactive (invisible)
Specific Reaction to G
Biosynthetic method
Producing various fragments
or
Terminated
Template
A
Juang RH (2004) BCbasics
3
Sangers Method How Terminated
5
ddNTP
Terminated
Normal Linking
Phosphodiester bond
3
dideoxynuceotide
Can not react
Juang RH (2004) BCbasics