Products restriction fragment analysis

Finally, restriction fragment analysis of PCR products can sometimes compensate for its inability to uncover all substitutions by allowing one to survey larger DNA segments (up to several kilobases in length). It must be noted that DGGE can be coupled with restriction fragment analysis to allow the detection of a fraction of the mutations that do not affect restriction sites.3... 

FIGURE 4.5 Analysis of mixtures of different meat species in processed and heated products. Restriction fragments of cytb amplicons (359 bp) obtained with Alu I (left) and Hae in (right) according to Meyer et al. (1995). Lanes 1 to 5 boiled sausages, 1 h, 70 to 75 C lane 1,1% beef, lamb, turkey, and chicken, each in pork lane 2,1% pork in beef lane 3,4% lamb, 48% pork, and 48% beef lane 4,1 % pork in turkey lane 5,1% pork in chicken. Lanes 6 to 9 preserved sausages [121°C, 3 to 5 (lanes 6 and 8)andE< 1 (lanes 7 and 9)] lanes 6 and 7,1% pork in beef lanes... 

Schwartz, H. E., Ulfelder, K., Sunzeri, F. J., Busch, M. R, and Brownlee, R. G., Analysis of DNA restriction fragments and polymerase chain reaction products towards detection of the AIDS (HIV-1) virus in blood, /. Chromatogr., 559, 267, 1991. 

Restriction endonucleases catalyze the hydrolysis of specific phosphodi-ester bonds in double-stranded DNA. These enzymes are often used to linearize a circular plasmid for hybrid DNA construction. They have also found use in the analysis of DNA and the construction of restriction maps. In this experiment, students will incubate various restriction enzymes with plasmid or viral DNA and analyze the product DNA fragments by agarose gel electrophoresis. 

Fig. 6. Physical detection of reciprocal translocations. The chromosomes shown are identical to those in Fig. S. Vertical arrows represent recognition sites for a single restriction enzyme horizontal arrows correspond to synthetic oligonucleotide primers and lines below the chromosomes indicate the sizes of restriction or PCR fragments. In (A), the alteration in restriction fragment size as a result of exchange is illustrated. Such alterations can be detected by Southern analysis using the duplicated sequence as a probe. In (B), the production of a PCR product from one of the exchange chromosomes is illustrated. Neither parental chromosome directs synthesis of a PCR product.

J. M. Butler, Separation of DNA restriction fragments and PCR products, m Analysis of Nucleic Acids by Capillary Electrophoresis (C. Heller, ed.), Verlag Vieweg, Wiesbaden, 1997, pp. 195-217. 

DNA applications, in particular DNA sequencing, human identification, and genetic analysis dominate the field. Other DNA applications including oligonucleotides, antisense DNA, restriction fragments, plasmids, PCR products, hydridization (DNA probe), and RNA have also been demonstrated [7,8]. 

In the laboratory. DNA can be cut into small pieces by enzymes called restriction endonucleases. The.se enzymes are restricted to cutting the DNA at specific base pair sequences. Tbe presence of polymorphic sites, where there are differences in the base pair sequence of the DNA, may create or abolish a cutting site for a particular restriction endonuclease. In these instances, the polymorphisms are known as restriction fragment length polymorphisms (Rf-LPs) as they lead to the production of DNA fragments of different lengths after enzyme action. DNA fragments may be separated by electrophoresis, transfered to a nylon membrane and hybridized with a DNA probe labelled with a radioactive or optically active marker. This is called. Southern blot analysis (Fig. 1). 

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