Kinetic PCR

Germer S, Holland MJ, Higuchi R. High-throughput SNP allele-frequency determination in pooled DNA samples by kinetic PCR. Genome Res Genome Res 2000 10(2) 258-266. 

Higuchi R, Fodder C, Dollinger G, Watson R. Kinetic PCR analysis real-time monitoring of DNA amplification reactions. Biotechnology (NY) 1993 11 1026-30. 

R. Higuchi, R. Watson, Kinetic PCR analysis using a CCD camera and without using oligo nucleotide probes. In PCR Applications (Eds. M.A. lnnis, D.H. Gelfand, J.J. Sninsky), Academic Press, San Diego, 1999, pp. 263-284. 

Ozawa, T., Tanaka, M., Ikebe, S., Ohno, K., Kondo, T. and Mizuno, Y. (1990) Quantitative determination of deleted mitochondrial DNA relative to normal DNA in parkinsonian striatum by a kinetic PCR analysis. Biochem. Bio-phys. Res. Commun. 172 483 89. 

Real-time (kinetic) PCR coupled to quantitation of productby SYBR Green 1 binding/ TaqMan M Microarrays... 

Real time PCR Sometimes referred to as kinetic PCR, real time PCR is a process by which a fluorescent signal is generated as the target sequence is amplified, and this fluorescent signal is measured at each cycle of amplification during the PCR process (Heid, 1996 Orlando et al, 1998). Two fundamental techniques are commonly used to generate the fluorescent signal ... 

Isolated RNA may contain tissue enzyme inhibitors and genomic DNA that result in reduced RT and PCR reaction efficiencies and generate unreliable and wrong quantification results. While this is not a problem for some applications, the tremendous amplification power of kinetic PCR may result in even the smallest amount of DNA contamination interfering with the desired specific amplification. To confirm the absence of residual DNA, a minus-RT should be included. Additionally it may be necessary to treat the RNA sample with commercially available RNAse-free DNAse, to get rid of residual DNA. Furthermore, the design of the PCR product should incorporate at least one exon to exon splice junction to allow a product obtained from the cDNA to be distinguished from genomic DNA contamination. 

More recently developed mathematical models with kinetic PCR efficiency correction allow for a more... 

Today, there is increasing appreciation of a more reliable normalization in relative quantification. It is vital to develop universal, artificial, and stable, internal standard materials that can be added prior to the RNA preparation to monitor the efficiency of RT as well as the kinetic PCR. Usually more than one housekeeping gene should be tested in a multiple... 

Higuchi, R., Fockler, C., Dollinger, G., et al., 1993. Kinetic PCR analysis real-time monitoring of DNA amplification reactions. Biotechnology 11, 1026-1030. Hu, G.K., Madore, S.J., Moldover, B., et al., 2001. Predicting splice variant from DNA chip expression data. Genome Res. 11, 1237-1245. 

Bubbles are formed instantaneously. This conclusion made in [33] is based on estimates taken from earlier works [37]. As seen from the above cited works by S. E. Sosin et al., this is not always true viscoelastic liquids under triaxial stretching stress are not destroyed instantly. The existence of an induction period may produce a considerable effect on foam growth kinetics upon free foaming, when pressure is lowered instantaneously from P > Pcr to P < Pcr in a melt with dissolved gas. However, it would appear that microfaults in polymer melts, which are caused by factors... 

Figure 2. Force generation and energy metabolism in human quadriceps femoris muscle stimulated intermittently at 20 Hz, with 1.6 sec tetani with 1.6 sec rest periods between tetani. The upper panel shows force, ATP turnover rate, and pH the middle panel, the concentrations of PCr, P and lactate and the lower panel, ATP, ADP, IMP, H, and calculated H2PO4. From Hultman et al. (1990), with permission from Human Kinetics Publishers.

The total amplification achieved by PCR is described by the expression, (1 + )", where E is the average per-cycle efficiency and n is the total number of cycles. The amount of target sequence and the variable presence of inhibitors in clinical specimens influence both the efficiency and the kinetics of amplification. As seen in the preceding expression, small differences in the efficiency of amplification are exponentially compounded and lead to very large and unpredictable differences in product yield. The situation is even more complicated when the target is RNA. PCR must be preceded by reverse transcription to produce complementary DNA (cDNA), and the efficiency of this process is another variable that may influence product yield. 

X. Ravalec, N. Le Tallec, F. Carre, J. D. de Certaines and E. Le Rumeur, Kinetics of PCr to ATP and beta-ATP to beta-ATP phosphoryl conversion are modified in working rat skeletal muscle after training. MAGMA, 1999, 9,52-58. 

Weinberger, . M. Wiedenmann, E. Bohm, S. Jilg, W. Sensitive and accurate quantitation of hepatitis virus DNA using a kinetic fluorescence detection system (TaqMan PCR). J Virol Methods 2000, 35(1-2), 75-82. 

Conventional gas chromatography (GC) based on the use of chiral stationary phases can handle only a few dozen ee determinations per day. In some instances GC can be modified so that, in optimal situations, about 700 exact ee and E determinations are possible per day [29]. Such meclium-throughputmay suffice in certain applications. The example concerns the lipase-catalyzed kinetic resolution of the chiral alcohol (R)- and (S)-18 with formation of the acylated forms (R)- and (S )-19. Thousands of mutants of the lipase from Pseudomonas aeruginosa were created by error-prone PCR for use as catalysts in the model reaction and were then screened for enantioselectivity [29]. 

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