An oligonucleotide microarray bait for isolation of target gene fragments.
A new molecular-baiting method was studied by retrieving targeted gene fragments from an oligonucleotide microarray bait after hybridization. To make the microarray bait, 70-mer oligonucleotides that were designed to specifically represent the SSA1 gene of Saccharomyces cerevisiae were printed on the slide. Samples of the Saccharomyces cerevisiae mRNA were extracted and labeled by the RD-PCR (Restriction Display PCR) method using the Cy5-labelled universal primer, then applied for hybridization. The sample fragments that hybridized to the microarray were stripped, and the eluted cDNAs were retrieved and cloned into the pMD 18-T vector for transformation, plasmid preparation, and sequencing. BLAST searching of the GenBank database identified the retrieved fragments as being identical to the SSA1 gene (from 2057-2541bp). A new method is being established that can retrieve the sample fragments using an oligo-microarray-bait.
Rong
Shi
S
Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, P.R. China.
Wen-li
Ma
M
Cui-hua
Liu
L
Yan-bin
Song
S
Xiang-ming
Mao
M
Wen-ling
Zheng
Z
D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics; D005456:Fluorescent Dyes; D018390:Gene Targeting; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics
D004269:DNA, Bacterial; D004271:DNA, Fungal; D005456:Fluorescent Dyes; D012333:RNA, Messenger