Novel methods for amplifying and detecting nucleic acid sequences

A process for non-linearly amplifying a specific nucleic acid sequence comprising the steps of:

• providing:
• said specific nucleic acid sequence,
• a first initial primer or a nucleic acid construct for said specific nucleic acid sequence, said first initial primer or nucleic acid construct comprising two segments:
• (A) a first segment (i) being substantially complementary to a first portion of said specific nucleic acid sequence and (ii) capable of template-dependent first extension, and
• (B) a second segment being (i) substantially non-identical to said first segment, (ii) substantially identical to a second portion of said specific nucleic acid sequence, (iii) capable of binding to a complementary sequence of said second segment and (iv) capable of providing for subsequent binding of a first segment of a second primer or nucleic acid construct to said first portion of said specific nucleic acid sequence under isostatic or limited cycling conditions, such that a second primer extension is produced to displace a first primer extension;
• and
• a subsequent initial primer or a nucleic acid construct to the complement of said specific nucleic acid sequence, said subsequent initial primer or nucleic acid construct comprising two segments,
• (A) a first segment (i) being substantially complementary to a first portion of said specific nucleic acid sequence and (ii) capable of template-dependent first extension, and
• (B) a second segment being (i) substantially non-identical to said first segment, (ii) substantially identical to a second portion of said specific nucleic acid sequence, (iii) capable of binding to a complementary sequence of said second segment and (iv) capable of providing for subsequent binding of a first segment of a subsequent primer to said first portion of said specific nucleic acid sequence under isostatic or limited cycling conditions, such that a second primer extension is produced and displaces a first primer extension;
• and substrates, buffer and a template-dependent polymerizing enzyme;
• and
• incubating said specific nucleic acid sequence and said novel primer or nucleic acid construct in the presence of said substrates, buffer and template-dependent polymerizing enzyme under isostatic or limited cycling conditions;
• thereby non-linearly amplifying said specific nucleic acid sequence.

A process for non-linearly amplifying a specific nucleic acid sequence comprising the steps of:

• providing:
• said specific nucleic acid sequence and its complement;
• a first initial primer or a nucleic acid construct for said specific nucleic acid sequence, said first initial primer or nucleic acid construct comprising two segments:
• (A) a first segment (i) being substantially complementary to a first portion of said specific nucleic acid sequence and (ii) capable of template-dependent first extension, and
• (B) a second segment being (i) substantially non-identical to said first segment, (ii) substantially identical to a second portion of said specific nucleic acid sequence, (iii) capable of binding to a complementary sequence of said second segment and (iv) capable of providing for subsequent binding of a first segment of a subsequent first primer to said first portion of said specific nucleic acid sequence under isostatic or limited cycling conditions, such that a second primer extension is produced and displaces said first primer extension;
• and
• a second initial primer or a nucleic acid construct complementary to said first primer extension, said second initial primer or nucleic acid construct comprising a segment characterized by capable of template-dependent extension under isostatic or limited cycling conditions;
• and
• substrates, buffer and a template-dependent polymerizing enzyme;
• incubating said specific nucleic acid sequence and said novel primer or nucleic acid construct in the presence of said substrates, buffer and template-dependent polymerizing enzyme under isostatic or limited cycling conditions;
• thereby non-linearly amplifying said specific nucleic acid sequence.

A process for non-linearly amplifying a specific nucleic acid sequence comprising the steps of:

• providing:
• said specific nucleic acid sequence;
• a singular primer or a singular nucleic acid construct capable of non-linear amplification, comprising three segments:
• (a) a first segment (i) being substantially complementary to a first portion of said specific nucleic acid sequence and (ii) capable of template-dependent first extension;
• (b) a second segment substantially identical to a second portion of said specific nucleic acid sequence;
• and
• (c) a third segment substantially identical to said first segment;
• wherein said first primer extension is capable of producing sequences that are capable of hybridizing to said second segment and is capable of self-priming and self-extending to produce a complement to said third segment, and
• substrates, buffer and a template-dependent polymerizing enzyme;
• and
• incubating said specific nucleic acid sequence and said primer or nucleic acid construct in the presence of said substrates, buffer and template-dependent polymerizing enzyme;
• thereby non-linearly amplifying said specific nucleic acid sequence.

A post-termination labeling process for nucleic acid sequencing comprising the steps of:

• producing, in the presence of untagged or unlabeled substrates, untagged or unlabeled primer, polymerizing enzyme, buffer and an appropriate untagged or unlabeled terminator for each nucleotide base, nucleic acid fragments corresponding to said nucleic acid sequence of interest, wherein each of said terminators comprise a chemically reactive group that covalently binds to a tagged molecule under conditions that internal sequences are substantially non-reactive to said tagged molecules and said chemical reactions do not substantially interfere with separation of said fragments in a medium or matrix;
• separating the fragments produced in a medium or matrix;
• and
• detecting said separated fragments by detecting said tagged molecule in said medium or matrix.