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5.10.6 Applications of the PCR Many traditional methods in molecular biology have now been superseded by the PCR and the applica ...

control DNA template amplified with separate primers at the same time as the specific target DNA. However these types of quantit ...

graph is prepared from which an unknown concentration can be deduced. Since SYBR green and similar DNA-binding dyes are non-spec ...

5.11 Nucleotide sequencing of DNA 5.11.1 Concepts of nucleic acid sequencing The determination of the order or sequence of bases ...

sequencing is by far the most popular method and many commercial kits are available for its use. However, there are certain occa ...

at cytosine, etc., it is possible to read the sequence of the newly synthesised strand from the autoradiogram, provided that the ...

1 ACGTACGT ACG Direction of electrophoretic movement T 23 Fig. 5.38 Autoradiograph of a DNA sequencing gel. Samples were prepare ...

It is also possible to undertake nucleotide sequencing from double-stranded molecules such as plasmid cloning vectors and PCR am ...

sequencing products. This is where liquid polymers in thin capillary tubes are used obviating the need to pour sequencing gels a ...

genome analysis a reality. Indeed more advanced methods using nanotechnology are in development and may provide an even more eff ...

separated by electrophoresis under denaturing conditions, and analysed separately. DNA labelled at one end is divided into four ...

6 Recombinant DNA and genetic analysis R. RAPLEY 6.1 Introduction 6.2 Constructing gene libraries 6.3 Cloning vectors 6.4 Hybrid ...

initial use the methods for the production of gene libraries have been steadily refined and developed. Although microarray analy ...

molecules, termedreannealing. All these pairings are transient, owing to the weak- ness of hydrogen bonding between the few base ...

a specialised DNA carrier molecule termed avector(Section 6.3). Thus each DNA fragment is inserted by ligation into the vector D ...

the nucleic acid starting material. Since the genome of an organism is fixed, chromo- somal DNA may be isolated from almost any ...

may need to be repaired to make them flush-ended. This can be achieved by using a modified DNA polymerase termed Klenow polymera ...

mRNA molecules the clones of each individual mRNA have to be synthesised. Libraries that represent the mRNA in a particular cell ...

the poly(A) of mRNA. Alkaline hydrolysis is then used to remove the RNA strand, leaving single-stranded DNA which can be used, l ...

many of the most commonly used restriction enzymes. Linkers are blunt-end ligated to the cDNA but since they are added much in e ...