Cartridge-based high-throughput purification of oligonucleotides for reliable oligonucleotide arrays.
A novel, cartridge-based procedure for the efficient and irreversible detritylation of oligonucleotides is reported. This method, combined with a process for the elimination of depurinated fragments produces, in a highly parallel fashion, oligonucleotides with better purity than those traditionally obtained using reversed-phase high-performance liquid chromotography purification. Our combined detritylation and purification methodology compares favorably with commercial cartridge-based purification systems. The benefits of working with pure oligonucleotides, with regard to higher signal and better signal linearity, are shown in array-based hybridization experiments.
Semenyuk
Andrey
A
Department of Genetics and Pathology, Uppsala University, 75185 Uppsala, Sweden.
Ahnfelt
Matilda
M
Estmer Nilsson
Camilla
C
Yong Hao
Xiao
X
Földesi
Andras
A
Kao
Yu-Shu
YS
Chen
Hong-Huei
HH
Kao
Wei-Chen
WC
Peck
Konan
K
Kwiatkowski
Marek
M
D057927:Hydrophobic and Hydrophilic Interactions; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000706:statistics & numerical data; D015345:Oligonucleotide Probes; D009841:Oligonucleotides / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results; D014320:Triphenylmethyl Compounds / Q000737:chemistry
D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D014320:Triphenylmethyl Compounds