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PubMed · 8053945

[Kinetic analysis for subtractive hybridization of transcripts].

E D Sverdlov·O D Ermolaeva
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Abstract

A theoretical analysis of kinetics of two subtractive cDNA hybridizations has been carried out: hybridisation of an excess of denatured double-stranded cDNA (driver) with denatured double-stranded cDNA (tracer) containing some sequences (targets) missing from the driver, and hybridisation of denatured double-stranded tracer with denatured double-stranded driver unable to be renatured. Our calculations show that the first strategy requires several rounds of hybridisation for sufficient enrichment of tracer with target sequences of the low-abundant mRNA: in the first round the renatured fraction is removed, and in subsequent rounds the remaining single-stranded fraction cDNA is used and renatured fraction is collected. The second strategy makes it possible to sufficiently enrich non-reassociated fraction of tracer with minor fractions of target sequences in a single round of subtractive hybridisation.

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DNA, ComplementaryKineticsModels, TheoreticalNucleic Acid DenaturationNucleic Acid HybridizationRNA, MessengerEnglish AbstractJournal Article

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E D Sverdlov, O D Ermolaeva. 1994. [Kinetic analysis for subtractive hybridization of transcripts].. https://pubmed.ncbi.nlm.nih.gov/8053945/

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