Amersham Pharmacia Biotech UK Limited

Nycomed Amersham Plc

The Radiochemical Centre Limited

Amersham Biosciences UK Limited

Alan L Hamilton

David Loakes

Adrian Christopher Simmonds

Clifford Leslie Smith

Daniel M Brown

Shiv R Kumar

Mark McDougall

Satyam Nampalli

Fergal Hill

William Jonathan Cummins

Mark Samuel Briggs

John Griffiths

John Mama

John C Charlton

Nigel Bosworth

Ian Edward Bruce

David Williams

Patrick Finn

Valerie Millar

Richard Martin West

Martyn N Birch

Brian Scott

Malcolm J Hatcher

Disclosed is a non-fluorescent cyanine dye that may be used as an acceptor in fluorescence energy transfer assays involving the detection of binding and/or cleavage events in reactions involving biological molecules, and assay methods utilising such dyes. The non-fluorescent cyanine dye is a compound of formula (I), wherein the linker group Q contains at least one double bond and forms a conjugated system with the rings containing X and Y; groups R , R , R and R are attached to the rings containing X and Y, or optionally, are attached to atoms of the Z and Z ring structures; Z and Z each represent a bond or the atoms necessary to complete one or two fused aromatic rings each ring having five or six atoms, selected from carbon atoms and, optionally, no more than two oxygen, nitrogen and sulphur atoms; at least one of groups R , R , R , R , R , R and R is a target bonding group; any remaining groups R , R , R , R and R groups are independently selected from the group consisting of hydrogen, C -C alkyl, OR , COOR , nitro, amino, acylamino, quaternary ammonium, phosphate sulphonate and sulphate, where R is selected from H and C -C alkyl; any remaining R and R are selected from C -C alkyl which may be unsubstituted or substituted with phenyl, the phenyl being optionally substituted by up to two substituents selected from carboxyl, sulphonate and nitro groups; characterised in that at least one of the groups R , R , R , R , R , R and R comprises a substituent which reduces the fluorescence emission of said dye such that it is essentially non-fluorescent.

Energy transfer assay method and reagent

Nucleoside analogues or base analogues having structure (I), where X&equals;O or NH or S, Y&equals;N or CHR or CR , W&equals;N or NR or CHR or CR or S, n&equals;1 or 2; each R is independently H or O or alkyl or alkenyl or alkoxy or aryl or a reporter moiety; where necessary (i.e. when Y and/or W is N or CR where R is not O) a double bond is present between Y and W or W and W, and Q is H or a sugar or a sugar analogue or a nucleic acid backbone or backbone analogue.

Base analogues

Nucleoside analogues have structure (2) wherein Q is H or a sugar moiety or sugar analogue or a modified sugar or a nucleic backbone or backbone analogue, W is an alkylene or alkenylene chain of 0-5 carbon atoms, any of which may carry a substituent R , X is O or N or NR or CR , X&prime; is O or S or N, provided that when X&prime; is O or S, then X is C, Y is CH or N, R is NH or SMe or SO Me or NHNH , each of R and R is independently H or F or alkyl or alkenyl or aryl or acyl or a reporter moiety, R is independently H or alkyl or alkenyl or aryl or acyl or a reporter moiety, and R is H or &boxH;O or F or alkyl or aryl or a reporter moiety.

Tricyclic base analogues

Dye intermediate and method

Nucleoside analogues containing the degenerate base analogue P and derivatives thereof are provided with reporter moieties preferably comprising signal moieties. The nucleoside analogues are useful for labeling DNA or RNA or for incorporating in oligonucleotides.

Nucleoside analogues

Nucleoside analogues in which a group M replaces the natural base where M is (1) or (2) or (3), where each of X,, X,and X,are C or N, each of R,and R,is the same or different and each is H, NO,, CO, COR,, OR,, CN, O, CON(R,),, COOR,, SO,R,, SO,R,, SR,, NHCHO, (CH,),(R,),, halogen, or a reporter moiety, each of R,and R,is H or hydrocarbyl or a reporter moiety, and n is 0-4. The analogues are substrates for polymerase and terminal transferase enzymes.

Squarate dyes and their use in fluorescent sequencing method

A method of making radioactively labelled vitamin B-12 by reacting .alpha.-(5,6-dimethylbenzimidazolyl)hydrogenobamide with cobaltous ion Co-57, or with another radioactive cobalt isotope. The method enables vitamin B-12 of high specific activity to be made, which is useful for in vitro and in vivo analysis.

Vitamin B-12 cobalt-57 and process

Growing community of inventors

Amersham, United Kingdom

Alan L Hamilton