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The Public Health Research Institute of the City of New York, Inc.

Alexander B Chetverin

Fred Russell Kramer

David Y Zhang

Ying Jie Wu

Helena Vladimirovna Chetverina

Oleg V Kurnasov

Viktor I Ugarov

Alexander S Spirin

Lyubov A Ryabova

Igor Y Morosov

Elena V Volianik

A method of sorting mixtures of nucleic acid strands comprising hybridizing the strands to an array of immobilized oligonucleotides, each of which includes a constant segment adjacent to a variable segment. The constant segment of the immobilized oligonucleotides can be made complementary to the ends of strands obtained by digesting a double-stranded nucleic acid with a restriction enzyme and restoring the restriction sites, thereby permitting the sorting of strands according to their variable sequences adjacent to their constant terminal restored restriction sites.

Method of sorting a mixture of nucleic acid strands on a binary array

Amplification and/or expression of nucleic acids is carried out in a medium immobilized by using an organic and/or inorganic solid matrix penetrating the medium and having a porous, fibrous, reticulated, coiled, capillary, lamellar or folded texture and which includes the components of a cell-free enzyme system of exponential amplification of nucleic acids and/or components of a cell-free enzyme system of nucleic acid expression. In this medium, the progeny of each molecule (clone) and the expression products remain in the same zone of the reaction volume where the matrix molecule was initially located. The method permits cloning of nucleic acids in vitro as well as detection of solitary nuleic acid molecules in the sample studied.

Method for amplification of nucleic acids in solid media

Batch and continuous-flow cell-free methods for synthesizing protein wherein translation is coupled with replication of recombinant mRNA by an RNA-directed RNA polyerase, such as Q.beta. replicase. Transcription of a DNA template to produce the recombinant mRNA can be coupled additionally with coupled transcription-replication-translation methods, both batch and continuous. The invention also includes kits of reagents for synthesizing user-selected proteins according to methods of this invention.

Coupled replication-translation methods and kits for protein synthesis

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Pushchino, Russia

Alexander B Chetverin