ROLL: a method of preparation of gene-specific oligonucleotide libraries.
The selection of nucleic acid sequences capable of specifically and efficiently hybridizing to target sequences is crucial to the success of many applications, including microarrays, PCR and other amplification procedures, antisense inhibition, ribozyme-mediated cleavage, and RNA interference (RNAi). Methods of selection using nucleotide sequence libraries have several advantages over rational approaches using defined sequences. However, the high complexity of completely random (degenerate) libraries and their high toxicity in cell-based assays make their use in many applications impractical. Gene-specific oligonucleotide libraries, which contain all possible sequences of a certain length occurring within a given gene, have much lower complexity and, thus, can significantly simplify and accelerate sequence screening. Here, we describe a new method for the preparation of gene-specific libraries using the ligation of randomized oligonucleotide probes hybridized adjacently on target polynucleotide templates followed by PCR amplification. We call this method random oligonucleotide ligated libraries (ROLL).
Vlassov
Alexander V
AV
SomaGenics, Inc., Santa Cruz, CA 95060, USA. avlassov@somagenics.com
Koval
Olga A
OA
Johnston
Brian H
BH
Kazakov
Sergei A
SA
D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000737:chemistry; D015723:Gene Library; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000235:genetics; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D012326:RNA Splicing; D012672:Semliki forest virus / Q000235:genetics
D018076:DNA, Complementary; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes