Methods and compositions for performing low background multiplex nucleic acid amplification reactions
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsJohan Erik Simon Fredriksson, Carl Oscar Fredrik Dahl
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Abstract
Methods and compositions for performing low background multiplex nucleic acid amplification reactions are provided. Aspects of the invention include contacting a nucleic acid sample with two or more primer pairs for two or more target nucleic acids under template dependent primer extension reaction conditions, e.g., polymerase chain reaction (PCR) conditions. The resultant amplified composition is then contacted with target nucleic acid circularizing reagents, and product circularized target nucleic acids are then selected, e.g., for further amplification. Also provided are systems and kits that find use in practicing embodiments of the inventions.
Claims24
A method of selectively amplifying two or more target nucleic acids of a nucleic acid sample, said method comprising:
- (a) contacting said nucleic acid sample with primer pairs specific for two or more target nucleic acid sequences of interest to produce a first reaction mixture;
- (b) subjecting said first reaction mixture to template-dependent primer extension reaction conditions to produce an amplified composition comprising target amplicons;
- (c) circularlizing said target amplicons in said amplified composition;
- and
- (d) selecting for said circularized target amplicons in said amplified composition.
(canceled)
17-18. (canceled)
A composition comprising two or more distinct circularized target amplicons, wherein each of said distinct circularized target amplicons is hybridized to its cognate circularization template nucleic acid.
(canceled)
A kit comprising:
- a set of two or more primer pairs specific for distinct target nucleic acid sequences;
- and
- a set of circularization template nucleic acids specific for the predicted target amplicons produced by each of said two or more primer pairs.
25-30. (canceled)
Dependencies are inferred from the claim text.