Amplification techniques

Walker, G. T., et al. (1992). Strand displacement amplification—an isothermal, in vitro DNA amplification technique. Nucleic Acids Res. 20, 1691-1696. 

Gutierrez M, Forster FI, McConnell SA, et al. The detection of CD2+, CD4+, CD8+, and WC1+ T lymphocytes, B cells and macrophages in fixed and paraffin embedded bovine tissue using a range of antigen recovery and signal amplification techniques. Vet. Immunol. Immunopathol. 1999 71 321-334. 

Rolling circle amplification (RCA) is an alternative method to polymerase chain reaction it is also a generic amplification technique that can be used in antibody assays. Using a replication process similar to that used by viruses, RCA allows the recognition,... 

T. Bacarese-Hamilton, L. Mezzasoma, C. Ingham, A. Ardizzoni, R. Rossi, F. Bistoni, and A. Crisanti, Detection of allergen-specific IgE on microarrays by use of signal amplification techniques. Clin. Chem. 48, 1367-1370 (2002). 

Alternative methods of diagnosis include enzyme immunoassay, DNA probes, and nucleic acid amplification techniques. 

The most widely used amplification technique is the polymerase chain reaction. Hardly a research study in molecular biology involving DNA amplification has been performed without using PCR. Many modifications of PCR have been described. Before we review these modified techniques, it would be appropriate to briefly review the PCR methodology. 

FIGURE 11.54 Diurnal profile of average (H02 + R02) concentrations measured at Cape Grim, Tasmania ( ), and at Mace Head, Ireland ( ), under clean air conditions using a chemical amplification technique. (Adapted from Carpenter el at., 1997.)... 

Subkhankulova T, Livesey FJ. Comparative evaluation of linear and exponential amplification techniques for expression profiling at the single-cell level. Genome Biol 2006 7 R18. 

Biochemical assays such as the dextran-coated charcoal (DCC) assay, certain signal amplification techniques, and other cytosol-based methods have been mostly replaced... 

Different authors and commercial suppliers have assigned different names to signal amplification using tyramine. For example, tyramine signal amplification (TSA) system and the catalyzed signal amplification (CSA) system are comma-daily available from DuPont NEN Life Science Products, Boston, MA, and DAKO Corporation, Carpinteria, CA, respectively. In addition, the tarns CARD (catalyzed reporter deposition) (Bobrow et al., 1989), TA (tyramide amplification) (Shindler and Roth, 1996), and ImmunoMax (Merz et al., 1995) have been used for the tyramine amplification technique. The use of different names for almost identical procedures has resulted in confusion. To standardize the terminology, the neutral abbreviation, tyramide amplification technique (TAT) should be accepted (Von Wasielewski et al., 1997). 

Antibody capture of viruses can be used as a preparatory step in nucleic acid amplification techniques. Immunocapture of virus particles can be used to streamline and/or optimize the concentration, purification and specificity requirements of polymerase chain reaction assays. 

Troutt A. B., McHeyzer-Williams M. G., Pulendran B. and Nossal G. J. (1992) Ligation-anchored PCR a simple amplification technique with single-sided specificity. Proc. Natl. Acad. Sci. USA 89, 9823-9825. 

The third amplification technique involved distinguishing chemical mutagenicity from other physiological processes. It was not uncommon in these promotional texts for scientist-activists to point out the levels of complexity that chemical mutagens present for the basic researcher, relative, for example, to radiation. In one text, Hollaender noted that there are only three or four types of radiation, but there are produced each year about 30,000 new compounds, of which probably 20 or 30 are mutagenic. 8 Other texts emphasized the physiological differences between mutagenicity and toxicity We must also remem-... 

The tyramide amplification technique is based on the ability of phenolic compounds to become oxidized to highly reactive and unstable intermediates (8). When biotinyl tyramide is oxidized, dimerization with electron-rich aromatic compounds, such as those found in protein molecules, occurs (9). This reaction can be harnessed in immunohistochemistry to generate highly reactive biotinyl-tyramide intermediates that bind rapidly to protein molecules in the immediate vicinity of peroxidase enzymes. This reaction results in the deposition... 

Random amplification of polymorphic DNA (RAPD) is an in vitro amplification technique (see section on polymerase chain reaction below) that eliminates the need for any prior characterization of the genome to be analyzed.12 14 Based on the random amplification of DNA segments using single short primers of arbitrary sequence, RAPD readily detects genetic polymorphisms in most organisms. The method is simple and rapid to... 

Indirect detection does require more steps, but oftentimes yields amplified signals relative to direct methods because layering of bridging molecules may increase the number of detector molecules per probe molecule. It is probably this bridging/amplification technique that has allowed current enzyme detection systems to approach the sensitivity of radiolabeled systems. The use of these indirect methods reduces steric problems that might arise from having enzyme molecules directly bound to probe molecules. 

Figure 1.14. Schematic diagram of the 1.56-pm pulse-generation system with the stimulated Raman seeding and amplification technique (Yamamoto et al., 2000). Reproduced with permission.

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