Nucleic Acids Research · 2000 · 136 citations · 10 references
EngineeringGeneticsMolecular BiologyMolecular GeneticsGenomicsFalse Positive ClonesSuppression Subtractive HybridizationMirror Orientation SelectionCloningMolecular DiversityBackground MoleculeMolecular Biological MethodOligonucleotideDna ReplicationGenetic VariationFunctional GenomicsBioinformaticsGenetic EngineeringSynthetic BiologySequence OrientationsSystems BiologyMedicineGenome Editing
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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Luda Diatchenko, Yun‐Fai Chris Lau, A. Patricia Campbell et al. · Proceedings of the National Academy of Sciences · 1996 · 3K citations · Full text
An improved PCR method for walking in uncloned genomic DNA
Paul D. Siebert, Alex Chenchik, D.E. Kellogg et al. · Nucleic Acids Research · 1995 · 1K citations · Full text
Chromatin, Dna Sequencing, Dna +13
A G-protein-coupled receptor for leukotriene B4 that mediates chemotaxis
Takehiko Yokomizo, Takashi Izumi, Kyungho Chang et al. · Nature · 1997 · 978 citations · Full text