Multiply-primed amplification of nucleic acid sequences
OwnerHaiguang Xiao, Scott Hamiltoninventors
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OwnerHaiguang Xiao, Scott Hamiltoninventors
Estimating the term…
Improved processes for the amplification of target DNA sequences in the form of single or double stranded 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 wherein some or all of the deoxyribonucleotides are replaced by deoxyribonucleotide analogues that reduce the Tm of the amplified product. The product of this amplification is used for DNA sequencing and other analyses that involve hybridization. Kits containing components for use in the invention is also described. Also described are further uses of this amplified DNA in sequencing, single base substitution detection, modifying the restriction enzyme fragmentation patterns and other molecular biology applications.
A method for amplifying nucleic acid sequences, comprising:
A kit for amplyfing nucleic acid sequences comprising multiple single stranded oligonucleotide primers, a DNA polymerase and one or more modified nucleoside triphosphates that upon incorporation changes the melting temperature (Tm) of amplified DNA products from the melting temperature (Tm) of said one or more amplification targets.
A kit for sequencing a nucleic acid comprising multiple single stranded oligonucleotide primers, a DNA polymerase and one or more modified nucleoside triphosphates that upon incorporation changes the melting temperature (Tm) of amplified DNA products from the melting temperature (Tm) of said one or more amplification targets, a DNA polymerase suitable for dideoxy chain-termination DNA sequencing, deoxynucleoside triphosphates, and at least one chain-terminating dideoxynucleoside triphosphate.
Dependencies are inferred from the claim text.