208
FRIEDHELM HILDEBRANDT AND IVA SINGH-SAWHNEY
A
reaion of jnterest
5 '
I
I
a)DNA
template
+ cccMCcGCn'f.CCCCCCCTTMTTMCCC
1111 111111 11111 11 111111111 1111
- GGGTTGGCCMGGGGGG~
b) primers
c) (primers
hybridized
+
to template)_
d)Taq.
polymerase
S'AACCGGTI
AATIAAT'l' S ,
CCCAACCGG CCCCCCCCTIAATTAACCC
111 II I II
~ 'AACCGGTT
AATIAATT 5'
II I II I II
GGGTTGGCCAAGGGGGGGGAATTAATTGGG
e) reaction buffer, dNTPs
8
D
+
5'
primer
5'
t 1. separate DNA strands
+
J. 2. hybridize oligonucleotide
c
f primers
~
i3
Temp .
(•C} denature
94
72
50
2'
exlond
3'
Time
(min.)
+ . . . . . . . . . . . . . ~ . . ~ ~
f!!i~
-II
J. 3. polymerize a new strand
l by primer extension
Fig. 1. Mechanisms of a polymerase chain reaction (PCR). A Components of a PCR; B Polymerase reaction; C Heat cycle for a PCR reaction; D A PCR cycle has 3 steps