Multiply-primed amplification of nucleic acid sequences
OwnerRoger S. Lasken, Frank B. Dean +1inventors
InventorsRoger S. Lasken, Frank B. Dean, John Nelson
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Abstract
Processes for the amplification of target DNA sequences in the form of single or double stranded circular DNA molecules, especially those present in colony and plaque extracts, using multiple specific and/or random sequence oligonucleotide primers are disclosed along with methods for detecting such amplified target sequences. A kit containing components for use in the invention is also described.
Claims68
A process for selectively amplifying nucleic acid sequences, comprising forming a mixture comprising:
- multiple single stranded non-circular oligonucleotide primers (P1), one or more amplification target circles (ATCs), a DNA polymerase and multiple deoxynucleoside triphosphates, under conditions wherein said ATC binds to more than one of said multiple P1 primers and wherein conditions promote replication of said amplification target circle by extension of the P1 primers to form multiple tandem sequence DNA (TS-DNA) products.
A process for selectively amplifying nucleic acid sequences, comprising:
- (a) mixing multiple single stranded non-circular oligonucleotide primers (P1) and one or more amplification target circles (ATC) under conditions wherein said ATC binds to one said multiple P1 primers to produce a primer-ATC sample mixture;
- (b) adding a DNA polymerase and multiple deoxynucleoside triphosphates under conditions that promote replication of said amplification target circle by extension of the P1 primers to form multiple primary tandem sequence DNA (TS-DNA) products.
A kit for amplifying DNA sequences comprising nuclease-resistant random primers, a DNA polymerase and one or more deoxyribonucleoside triphosphates.
A kit for amplifying DNA sequences comprising both nuclease-sensitive and nuclease-resistant random primers, a DNA polymerase and one or more deoxyribonucleoside triphosphates.
Dependencies are inferred from the claim text.