Nucleic Acids in Chemistry and Biology
5.1 DNA Sequence Determination 5.1.1 Principles of DNA Sequencing There are two major ways of determining the sequence of a DNA ...
Nucleic Acids in Biotechnology 169 Figure 5.1 The principles of classical DNA sequencing. A primer oligodeoxynucleotide is annea ...
each with a different dye attached (Section 8.5.4, Figure 8.11). The four sequencing reactions are carried out together in one r ...
namely a prokaryotic origin of DNA replication in the case of bacterial artificial chromosomes(BACs) or a centromere(the region ...
In some cases, nucleic acid hybridisation(annealing of complementary strands, Section 5.5) is useful to find the desired clone. ...
Nucleic Acids in Biotechnology 173 Figure 5.4 Transposon tagging 5.2.1.6 Chromosome Walking. If an overlapping series of clones ...
The PCR method has also found widespread application in forensics for the detection and amplification of very tiny amounts of DN ...
Type I consists of a large enzyme complex containing subunits encoding endonuclease, methylase and several other activities. The ...
5.3.3 Polynucleotide Kinase A polynucleotide kinase isolated from bacteriophage T4 catalyses the transfer of the -phosphate of ...
it will also catalyse the joining of two oligoribonucleotides in the presence of a complementary splint oligodeoxyribonucleotide ...
Other features of the synthetic gene are internal restriction enzyme sites (shaded), which can be intro- duced artificially mere ...
encoded by a gene and to determine its levels of transcription and tissue specificity. Methods exist for both of these purposes ...
factors. At low temperatures, the re-association rate is determined by the difference in free energy between the unassociated an ...
concentration of the corresponding gene in the sample. This latter technique is called dot blottingand is very useful in this li ...
number of stutters. The problem was how to access them. Jeffreys observed that a chance-studied min- isatellite tucked away insi ...
5.5.3.6 PCR-Based Methods. The discovery of short tandem repeats(STRs) together with the introduction of automated sequencing te ...
and laboratory information management systems, including bar coding of samples, to reduce operator errors. A typical electropher ...
of a test and a standard probe provides an immediate signal of the presence or absence of a particular sequence. Ultimately, suc ...
incubated with the surface and chemical coupling occurs only at those sites that have been irradiated in the preceding step. Lig ...
build oligodeoxyribonucleotides on glass surfaces. An entirely new carbonate-based protecting group chem- istry enables deprotec ...
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