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AssigneeThe Secretary of State for Defence
InventorsDavid James Squirrell, Martin Alan Lee
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A method of analysing the length of a particular region within a nucleic acid, said method comprising a) subjecting a sample of said nucleic acid to a plurality of amplification reactions in which said region is amplified, wherein the time of the extension phase in each of said reactions is varied; b) monitoring the progress of said amplification reactions; c) determining the minimum time during which extension phase of the amplification is completed within each reaction mixture and relating that to the length of the sequence undergoing extension. The method, combined with melting point analysis will allow percentage GC content of a sequence to be determined. Length analysis of this type can be used in diagnosis or analysis as well as in recombinant DNA technology to check for the presence of concatamers, and in taxonomic classification or forensics. Apparatus for use in the method is also described and claimed.
A method of analysing the length of a particular region within a nucleic acid, said method comprising
A method according to any one of the preceding claims wherein the said plurality of amplification reactions are effected simultaneously.
A method according to any one of the preceding claims wherein each amplification reaction is effected in a well of a multiwell vessel, and each well is heated by supplying current to an electrically conducting polymer which comprises or is able to heat said well.
A method according to claim 6 wherein the time of the extension phase of the amplification reaction in each well is independently controllable.
A method according to any one of the preceding claims wherein the progress of the amplification reaction is monitored using a fluoresent label means, present in the reaction.
A method according to any one of the preceding claims wherein in step (b), the progress of the extension phase is monitored continuously.
A method according to any one of the preceding claims which is to screen clones obtained by recombinant DNA technology for the presence of concatemers.
A method according to any one of the preceding claims wherein the temperature at which particular reaction products form duplexes or duplex forms destabilise within said sample is noted, and this information, combined with the determination of the length of the sequence, is used to calculate the percentage GC content of the sequence.
Apparatus for use in a method according to any one of the preceding claims.
Dependencies are inferred from the claim text.