MULTIPLY-PRIMED AMPLIFICATION OF NUCLEIC ACID SEQUENCES

摘要

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.