Multiplex amplification of polynucleotides
AssigneeApplied Biosystems, LLC
InventorsMark ANDERSEN, David RUFF
Life of this patent
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Abstract
The present invention provides methods, reagents and kits for carrying out a variety of assays suitable for analyzing polynucleotides or samples that include an amplification step performed in a multiplex fashion. Also provided are methods for analyzing and improving the efficiency of amplification and for carrying out gene expression analysis.
Claims21
1-56. (canceled)
A method of simultaneously amplifying a plurality of polynucleotide sequences of interest in a single reaction comprising:
- a) selecting two or more polynucleotide sequences of interest for amplification;
- b) hybridizing to the polynucleotide sequences of interest a primer set, wherein each primer set contains at least one uracil nucleobase;
- and
- c) simultaneously amplifying the polynucleotide sequences of interest by polymerase chain reaction in a single reaction to generate a plurality of amplicons.
A method of generating a plurality of target sequences of interest, comprising the step of:
- amplifying by polymerase chain reaction one or more target polynucleotides in the presence of a plurality of amplification primers suitable for amplifying the target sequences of interest, wherein substantially all the amplification primers comprise at least one uracil nucleobase.
A kit for amplifying multiple polynucleotide sequences of interest in a single reaction comprising, a plurality of amplification primer sets suitable for amplifying the polynucleotide sequences of interest, wherein each primer set contains at least one uracil nucleobase, a thermostable DNA polymerase and a mixture of nucleotide triphosphates suitable for extension of the amplification primers via template dependent DNA synthesis.
Dependencies are inferred from the claim text.