A calibrated diversity assay for nucleic acid libraries using DiStRO--a Diversity Standard of Random Oligonucleotides.
We have determined diversities exceeding 10(12) different sequences in an annealing and melting assay using synthetic randomized oligonucleotides as a standard. For such high diversities, the annealing kinetics differ from those observed for low diversities, favouring the remelting curve after annealing as the best indicator of complexity. Direct comparisons of nucleic acid pools obtained from an aptamer selection demonstrate that even highly complex populations can be evaluated by using DiStRO, without the need of complicated calculations.
Schütze
Tatjana
T
Institute for Chemistry/Biochemistry, Free University Berlin, Thielallee 63, Berlin, Germany.
Arndt
Peter F
PF
Menger
Marcus
M
Wochner
Aniela
A
Vingron
Martin
M
Erdmann
Volker A
VA
Lehrach
Hans
H
Kaps
Christian
C
Glökler
Jörn
J
D002138:Calibration; D004247:DNA / Q000592:standards; D015723:Gene Library; D007700:Kinetics; D009691:Nucleic Acid Denaturation; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000592:standards*; D012015:Reference Standards; D052156:SELEX Aptamer Technique; D013696:Temperature
D009838:Oligodeoxyribonucleotides; D004247:DNA