Methods in Molecular Biology • 16 Enzymes of Molecular Biology

(Nancy Kaufman) #1
Taq Polymerase 35

Cydel

Cyde2

Cycle 3

t Dan~ure and anneal pdmere
m m
t Pdmer e0dention

t Danalure and anneal pdmers
emm m

m m
t Primer extention

t Denature and anneal pdmers
m m

m wm
mm me

m m m

t Palmer extention
umm

m
m

Fig. 1. Principle of the polymerase chain reaction.

amplification. Therefore, this system offers an optimal ratio between
specific and unspecific signal.
Characteristically, <1 lag of template in a vol of 100 ilL is used in the
PCR. In this experimental outline, a single copy gene of the human
genome would be in a concentration of 2.5 fM. Primers and enzyme are
used in large excess (1 pM and 1 nM, respectively). The efficiency of

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