Polymerase chain reaction amplified DNA

Tunis. M.A., K.B. Myambo, D.H. Gelfand and M.A. Brow 1988. DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA. Proc. Natl. Acad. Sci. USA 85 9436-9440. 

E. Kawasaki, R. Saiki, H. Erlich, Genetic Analysis Using Polymerase Chain Reaction-amplified DNA and Immobilized Oligonucleotide Probes Reverse Dot-Blot Typing , Meth. Enzymol., 218, 369-381 (1993). 

Sequencing of Polymerase Chain Reaction-Amplified DNAs... 

McCord BR, McClure DL, Jung JM (1993) Capillary electrophoresis of polymerase chain reaction-amplified DNA using fluorescence detection with an intercalating dye. J Chromatogr 652 75-82. 

Srinivasan K, Girard JE, Williams P, Roby RK, Weedn VW, Morris SC, Kline MC, Reeder DJ (1993a) Electrophoretic separations of polymerase chain reaction-amplified DNA fragments in DNA typing using a capillary electrophoresis-laser induced fluorescence system. / Chromatogr 652 83-91. 

Dl. Dean, D., Pant, C. R., and O Hanley, P., Improved sensitivity of a modified polymerase chain reaction amplified DNA probe in comparison with serial tissue culture passage for detection of Chlamydia trachomatis in conjunctival specimens from Nepal. Diagn. Microbiol. Infect. Dis. 12, 133-137 (1989). 

LaPointe, G., Leblanc, D. Morin, A., Use of a polymerase-chain-reaction-amplified DNA probe from Pseudomonas putida to detect D-hydantoinase-producing microorganisms by direct colony hybridization. Appl. Microbiol. Biotechnol., 42 (1995) 895-900. 

Doktycz, M.J., Hmst, G.B., Habibi-Goudarzi, S., McLuckey, S.A., Tang, K., Chen, C.H., Uziel, M., Jacobson, K.B., Woychik, R.P., and Buchanan, M.V. (1995) Analysis of polymerase chain reaction-amplified DNA products by mass spectrometry using matrix-assisted laser desorption and electrospray current status. Anal. Biochem., 230 (2), 205-214. 

The Polymerase Chain Reaction Amplifies Specific DNA Sequences... 

Nuovo, G. J., MacConnell, P. B, Simsir, A., Valea, F, and French, D. L. (1995) Correlation of the in situ detection of polymerase chain reaction-amplified metalloproteinase complementary DNAs and their inhibitors with prognosis m cervical carcinoma. Cancer Res. 55,267—275. 

Jensen MA, Webster JA, Straus N (1993) Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms. Appl Environ Microbiol 59 945-952... 

In conclusion, this integrated platform allows the synthesis of enzymes inside microdroplets through IVTT, monitoring of the enzyme activity in each of the droplets, and selection of the droplets with high enzyme activity. In conjunction with other molecular methods such as polymerase chain reaction, the DNA in these high-yielding droplets can be amplified individually and fiuther analyzed. 

CR Polymerase Chain Reaction. Widely used method for amplifying a DNA base sequence... 

The plaque assay is desirable because it is very sensitive and only detects infectious viral particles. However, there are viral agents which cannot be supported by cell lines. In these cases other methods must be used. The polymerase chain reaction (PGR), which amplifies DNA or RNA from viral agents, can be used to detect the presence and quantity of viral agents. The amount of RNA or DNA target in the initial sample can be determined by competitive PGR where the quantity of amplified product is compared to a control PGR product where the initial amount of target is known. Quantification is also possible by an end-point dilution method similar to that used to determine a tissue culture infections dose. PGR methods can be very sensitive however. 

I been established to serve as a registry of convicted offenders. When a DNA sample is obtained from a crime scene, the sample is subjected to cleavage with restriction endonucleases to cut out fragments containing the STR loci, the fragments are amplified using the polymerase chain reaction, and the sequences of the fragments are determined. 

Sequencing of DNA is carried out by the Sanger dideoxy method, and small DNA segments can be synthesized in the laboratory by automated instruments. Small amounts of DNA can be amplified by factors of 106 using the polymerase chain reaction (PCR). 

Polymerase chain reaction, PCR (Section 28.8) A method for amplifying small amounts of DNA to produce larger amounts. 

Differential display is a method for identifying differentially expressed genes, using anchored oligo-dT, random oligonucleotide primers and polymerase chain reaction on reverse-transcribed RNA from different cell populations. The amplified complementary DNAs are displayed and comparisons are drawn between the different cell populations. 

While many diseases have long been known to result from alterations in an individual s DNA, tools for the detection of genetic mutations have only recently become widely available. These techniques rely upon the catalytic efficiency and specificity of enzyme catalysts. For example, the polymerase chain reaction (PCR) relies upon the ability of enzymes to serve as catalytic amplifiers to analyze the DNA present in biologic and forensic samples. In the PCR technique, a thermostable DNA polymerase, directed by appropriate oligonucleotide primers, produces thousands of copies of a sample of DNA that was present initially at levels too low for direct detection. 

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.

Molecular methods used to uncover mutations are subject to several variables. The anticoagulants used for blood collection can affect digestion with restriction enzymes and amplification reactions. The type of detergent used in cell lysis can affect amplification of DNA by inhibiting the DNA-amplifying enzyme such as the taq polymerase used in the polymerase chain reaction (116). The control of contamination is crucial in ensuring the quality of results obtained by molecular analysis (117). 

Polymerase chain reaction (PCR) is one of the most important techniques for rapid bacterial identification. It consists of repeated cycles of enzymatic reactions in a thermal cycler (PCR machine) that copies DNA strands many times. The DNA amplified in one PCR cycle is used as a template for the next cycle. This results in an exponential increase of the desired target... 

Naito, Y. Ishikawa, K. Koga, Y. Tsuneyoshi, T. Terunuma, H. Arakawa, R. Molecular mass measurement of polymerase chain reaction products amplified from human blood DNA by electrospray ionization mass spectrometry. Rapid Commun. Mass Spectrom. 1995,9,1484-1486. 

Guo, Z.G. and Johnson, A.M. (1995) Genetic characterization of Toxoplasma gondii strains by random amplified polymorphic DNA polymerase chain reaction. Parasitology 111, 127-132. 

Joachim, A., Daugschies, A., Christensen, C.M., Bj0rn, H. and Nansen, P. (1997) Use of random amplified polymorphic DNA-polymerase chain reaction... 

Wu, Z., Nagano, I. and Takahashi, Y. (1998) The detection of Trichinella with polymerase chain reaction (PCR) primers constructed using sequences of random amplified polymorphic DNA (RAPD) or sequences of complementary DNA encoding excretory-secretory (E-S) glycoproteins. Parasitology 117, 173-183. 

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