Endonucleases, polymerase chain

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. 

Baumann, R. E., and Henschen, A. H. (1993). Human fibrinogen polymorphic site analysis by restriction endonuclease digestion and allele-specific polymerase chain reaction amplification Identification of polymorphisms at positions A alpha 312 and B beta 448. Blood 82, 2117-2124. 

A cassette-replacement approach was used to facilitate the introduction of amino acid mutations at various sites of the thrombin receptor. First, unique endonuclease restriction enzyme sites were generated at several positions within the thrombin receptor cDNA by mutating the nucleotide sequences. Second, the polymerase chain reaction (PCR) with primers encoding for the desired mutations was used to generate the cDNA cassette with the appropriate endonuclease restriction enzyme sites for replacement of the wild-type sequence. The locations for the introduction of the sites were chosen based on two requirements. They needed to be at or near regions of the cDNA sequence that codes for amino acids at junctions of transmembrane domains and extracellular loops. Also, introduction of the sites did not alter the amino acid sequence of the protein. The site-directed mutagenesis method of Kunkel et al.28 was used to introduce the mutations required for generating the... 

DNA sequencing Maxam-Gilbert method restriction endonuclease restriction fragment palindrome Sanger dideoxy method DNA synthesis DMT ether phosphoramidite phosphite polymerase chain reaction (PCR)... 

Restriction endonucleases and DNA ligase Cutting, rejoining DNA Reverse transcriptase Conversion of mRNA into cDNA Chemical DNA synthesis Oligonucleotides Polymerase chain reaction (PCR)... 

The technology for studying DNA involves looking for the differences in the fragments formed when the DNA is digested by specific enzymes known as restriction endonucleases, or amplification of selected regions of the DNA by a technique called the polymerase chain reaction. 

Loureiro, 2000 Deak, 2002 Capece et al., 2003). Each requires initial digestion of the harvested DNA using restriction endonucleases, enzymes that cleave DNA at specific nucleotide sequences unique for that enzyme. The DNA digest is the amplified at specific or randomly selected regions by polymerase chain reaction (PCR). Fragments are subsequently separated electrophoretically and their patterns compared against those of other isolates or databases. 

Exonuclease activity In addition to having the 5 —>3 po ) merase activity that synthesizes DNA, and the 3 ->5 exonucleas activity that proofreads the newly synthesized DNA chain lik DNA polymerase III, DNA polymerase I also has a 5 - 3 exon clease activity that is able to hydrolytically remove the RN primer. [Note These activities are exonucleases because the remove one nucleotide at a time from the end of the DNA chaii rather than cleaving it internally as do the endonucleases (Figui 29.18).] First, DNA polymerase I locates the space ("nick between the 3 -end of the DNA newly synthesized by DNA pol] merase III and the 5 -end of the adjacent RNA primer. Next, DN... 

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