posted on 2019-11-12, 17:33authored byJeewon Lee, Sunghoon Heo, Duhee Bang
Recombinase
polymerase amplification (RPA) is an isothermal DNA
amplification method with broad applications as a point-of-care test
and in molecular biology techniques. Currently, most of the applications
are focused on target-specific amplification. Because RPA has the
advantage of amplifying DNA under isothermal conditions, we utilized
RPA as a DNA library amplification tool. In this study, we used a
sheared genomic DNA library and an oligonucleotide (oligo) library
for the comparison of polymerase chain reaction and RPA. For the sheared
DNA library, we observed biased amplification after RPA was conducted.
Thus, to amplify the size-variable DNA library uniformly, we introduced
a linear amplification strategy with RPA and successfully improved
the uniformity. On the other hand, using the same-sized oligo library,
we confirmed that RPA amplified this library uniformly without modification
of the protocol. These results demonstrate that RPA can be applied
not only to amplify a specific target as previously demonstrated but
also to amplify a complex DNA library composed of a large number of
different DNA molecules.