Mirror orientation selection (MOS): a method for eliminating false positive clones from libraries generated by suppression subtractive hybridization | Zendy

Denis Rebrikov | Zendy

Open Access

Mirror orientation selection (MOS): a method for eliminating false positive clones from libraries generated by suppression subtractive hybridization

Author(s) -

Denis Rebrikov

Publication year - 2000

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/28.20.e90

Subject(s) - suppression subtractive hybridization , biology , adapter (computing) , complementary dna , genetics , orientation (vector space) , computational biology , microbiology and biotechnology , nucleic acid thermodynamics , cdna library , base sequence , gene , geometry , mathematics , electrical engineering , engineering

Suppression subtractive hybridization (SSH) is one of the most powerful and popular methods for isolating differentially expressed transcripts. However, SSH-generated libraries typically contain some background clones representing non-differentially expressed transcripts. To overcome this problem we developed a simple procedure that substantially decreases the number of background clones. This method is based on the following difference between target and background cDNAs: each kind of background molecule has only one orientation with respect to the two different flanking adapter sequences used in SSH, while truly differentially expressed target cDNA fragments are represented by both sequence orientations. The described method selects the molecules that arose due to hybridization of such mirror-orientated molecules. The efficiency of this method was demonstrated in both model and real experimental subtractions.

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