Polymerase chain reaction product sequencing

Greiner TC, Raffeld M, Lutz C, Dick F, Jaffe ES. Analysis of T cell receptor-gamma gene rearrangements by denaturing gradient gel electrophoresis of GC-clamped polymerase chain reaction products. Correlation with tumor-specific sequences. Am J Pathol 1995 146 46-55. 

Sebastio, P., Zanelli, P., andNeri, T.M. 2001. Identification of anchovy Engraulis encrasicholus L. and gilt sardine Sardinella aurita by polymerase chain reaction, sequence of their mitochondrial cytochrome b gene, and restriction analysis of polymerase chain reaction products in semipreserves. J. Agric. Food Chem. 49, 1194-1199. 

Muhammad WT, Tabb DL, Fox KF, and Fox A (2003) Automated discrimination of polymerase chain reaction products with closely related sequences by software-based detection of characteristic peaks in product ion spectra. Rapid Communications in Mass Spectrometry 16 1-8. 

Figure 40-7. The polymerase chain reaction is used to amplify specific gene sequences. Double-stranded DNA is heated to separate it into individual strands. These bind two distinct primers that are directed at specific sequences on opposite strands and that define the segment to be amplified. DNA polymerase extends the primers in each direction and synthesizes two strands complementary to the original two. This cycle is repeated several times, giving an amplified product of defined length and sequence. Note that the two primers are present in excess.

Fig. 9.4 Establishment of the quantitative methylation-specific polymerase chain reaction (MSP) analytical procedure. The ABI PRISM 7900 HT Sequence Detector system was used to perform real-time polymerase chain reaction (PCR) using MSP primers and bisulfite-modified template DNA. Upper panels Setting up the conditions to obtain the standard curves with 50% (A) or 25% (B) sequential dilution of the template. Lower panels The amplification curves on the left represent P-actin, unmethylated, and methylated MSP products, respectively, for reelin (RELN) (C). Amplification curves were compared at the set threshold before 40 cycles. Amplification curves from various samples are shown in the lower panel right (D)...

Are there any other possible uses for the construction of complex topological species One possible application is in the mass production of DNA polyhedral catenanes by biological means, such as the polymerase chain reaction (PCR) (Saiki et al. 1986) or by production in vivo. Figure 21 illustrates that semi-conservative replication (the mechanism used by DNA polymerases) cannot reproduce a stable branch. The DNA with different sequences in the two arms of the branch (cartooned as dashed and solid lines) leads to two heterologous duplex DNA molecules, rather than a second branched molecule. 

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