遗传学实验技术

In this chapter, we introduce our developed freeware tool Methyl-Typing. It provides methylation-related bioinformatics with a special focus on combined bisulfite restriction analysis (COBRA). We give an overview of the implementation and program modules for Methyl-Typing. Va ...

Closed-tube PCR methods (sometimes referred to as in-tube PCR methods) for locus-specific DNA �methylation analysis are methodologies in which the amplification and analysis of bisulphite-modified DNA take place in one tube without the need to remove the PCR products for further anal ...

Bisulphite pyrosequencing is a quantitative methodology for the investigation of DNA methylation of sequences up to 100-bp in length. Biotin-labelled, single-stranded PCR products generated from bisulphite-treated DNA are used as a template with an internal primer to perform the p ...

Phylogenetic diagrams (“trees of life”) based on computer-generated analyses of nucleic acid (DNA, RNA) or protein (amino acid residues) sequences are purported to reconstruct evolutionary history of the living organisms from which the macromolecules were isolated (1). “Horizon ...

Horizontal gene and genome transfer forces us to recognize that life evolves by fusion as well as bifurcation of lineages, and necessitates the expansion of traditional views of evolution. This chapter reviews the role that horizontal gene transfer (HGT) may play in integrating selection ...

This chapter defines the agents that provide for the movement of genetic material which fuels the adaptive potential of life on our planet. The chapter has been structured to be broadly comprehensive, arbitrarily categorizing the mobilome into four classes: (1) transposons, (2) plasmids, ...

Bacteria experience recombination in two ways. In the context of the Biological Species concept, allelic exchange purges genic variability within bacterial populations as gene exchange mediates selective sweeps. In contrast, horizontal gene transfer (HGT) increases the size of ...

We describe the reasons why the newly recognized process of horizontal gene transfer (HGT) forces evolutionists who study classification and microbiology to go beyond the classical Darwinian framework. We recall the importance of processes in philosophical definitions of speci ...

The success of proteomics relies heavily in the ability to characterize very diverse species of proteins. This diversity stems from a proteins physicochemical properties, its copy number and abundance and its association with other proteins. Prefractionation and simplificati ...

Current methods for quantitatively comparing complex protein profiles such as two-dimensional gel electrophoresis (2-DE), 2-D differential in-gel electrophoresis (DIGE), and liquid chromatography (LC)-mass spectrometry (MS) have limited resolution and dynamic ranges ...

Protein fractionation is essential to uncovering low-abundance proteins in complex protein mixtures. Many common methods and techniques are used to fractionate proteins, including chromatography (size exclusion, affinity, ion exchange, etc.), electrophoresis, and solut ...

End-labeling is a rapid and sensitive method for radioactively, or nonisotopically, labeling DNA fragments and is useful for visualizing small amounts of DNA. End-labeling can also be used to label fragments at one end. All of the enzymes employed are specific to either the 3′ or 5′ termini of DNA and wil ...

One of the most common precursors to undertaking a protocol for mutation detection is the production of a suitably labeled DNA probe (1). Labeled nucleotides (radioactive or fluorescent) can be incorporated efficiently into doublestranded DNA by a number of methods. One of the most common is by a ...

Electrophoresis through agarose or polyacrylamide gels is a standard method used to separate, identify, and purify nucleic acids. The technique is simple, rapid to perform and capable of resolving fragments that differ by as little as 0.2% in size. Electrophoresis occurs under the influen ...

Caspases are cysteine-aspartic acid specific proteases that are activated in response to different cell death inducing stimuli (1–3). Their activation initiates specific cleavage of the respective target proteins and therefore is considered to be a marker of the irreversible steps ...

p53 plays a central role in the cellular response to DNA damage. Induction of this protein is triggered by DNA breaks and correlates with their presence (1,2). In addition, p53 is upregulated by a number of different stress conditions including hypoxia, oncogene activation, viral infection, and ...

This chapter presents a new method for the detection of DNA damage that leads to the physiological process of apoptosis which is a normal programmed cell-death response to sufficiently toxic levels of DNA damage. The high-frequency ultrasonic detection of programmed cell death has been de ...

Poly (ADP-ribose) polymerase (PARP-1; EC 2.4.2.30) is a predominantly nuclear enzyme long presumed to function in the identification and repair of DNA strand nicks and breaks (for recent reviews see: 1–3). PARP is comprised of a N-terminal DNA-binding domain, a central automodification doma ...

DNA sequencing is the gold standard in DNA diagnostics and is the only absolute means of establishing the identity of a new mutation. However, the clinical cost of obtaining this information is often prohibitive, particularly when large DNA fragments are interrogated for the presence of any of a ...

Single-strand conformation (SSCP) and heteroduplex analysis are separate mutation scanning methods in their own right. They are, however, unusual in that they can be carried out simultaneously on a single gel.