基因表达差异显示实验

Matrix-assisted laser desorption time-of-flight mass spectrometry coupled with allelespecific primer extension is a proven method for typing single nucleotide polymorphisms (SNPs). A novel modification upon this methodology is the incorporation of a photocleavable lin ...

In forensic casework analysis, mitochondrial deoxyribonucleic acid (DNA) often is used when the evidence material contains scarce amounts of DNA. Here, a mitochondrial DNA typing system for D-loop and coding region analysis based on pyrosequencing is described. Pyrosequencing is a r ...

We describe a rapid and robust assay to genotype mitochondrial deoxyribonucleic acid (mtDNA) coding region single nucleotide polymorphism (SNPs) using the SNaPshot (Applied Biosystems, Foster City, CA) minisequencing reaction kit. A protocol for mtDNA SNaPshot typing is descri ...

We describe a method for the simultaneous typing of Y-chromosome single nucleotide polymorphism (SNP) markers by means of multiplex polymerase chain reaction (PCR) strategies that allow the detection of 35 Y chromosome SNPs on 25 amplicons from 100 to 200 pg of chromosomal deoxyribonucl ...

Single nucleotide polymorphisms (SNPs) are the most frequent polymorphisms described in the human genome, and their analysis is becoming an extensive routine in molecular biology, not only in the forensic field, but also in population and clinical genetics. In particular, SNPs located on ...

Genetic polymorphisms of genes coding for metabolic enzymes are helpful to predict how an individual may respond to medication or drugs. The described approach for the identification of genetic variations for the cytochrome P450 enzymes CYP2D6 and CYP2C19 has been designed for the rapid ...

Whereas the analysis of ancient DNA (aDNA) has become an increasingly popular mode of investigation in both archaeological and evolutionary studies, this approach is complicated by the degraded nature of ancient nucleic acids, the presence of enzymatic inhibitors in aDNA extracts, as w ...

Molecular analysis of fossil and archaeological remains has been established as a powerful tool in providing new insight in phylogenetic investigations. The overlapping set of molecular modifications and degradation that forensic samples share with archaeological specim ...

Forensic laboratories are increasingly confronted with problematic samples from the scene of crime, containing only minute amounts of deoxyribonucleic acid (DNA), which may include polymerase chain reaction (PCR)-inhibiting substances. Efficient DNA extraction proced ...

Since the discovery of deoxyribonucleic acid (DNA) profiling in 1985, forensic genetics has experienced a continuous technical revolution, both in the type of DNA markers used and in the methodologies or its detection. Highly informative and robust DNAtyping systems have been develop ...

We present a fluorogenic real-time polymerase chain reaction (PCR) procedure to target a segment (106-112 base pairs ) of the X-Y homologous amelogenin gene by measuring the 5′' nuclease activity of the Taq deoxyribonucleic acid (DNA) polymerase using two (X-Fam-labeled and Y-Vic-labeled) ...

Many large mammalian species are on the verge of extinction in part because of the trade in their skin, bone, horn, or body parts for supposed medicinal purposes. Identification of the species is required to determine that a crime has been committed. This chapter details a robust DNA technique using p ...

Alteration of genomically encoded nucleotide sequence in mRNA is called RNA editing. RNA editing was first described in mitochondrial mRNAs of kinetoplastid protozoa (Trypanosoma, Leishmania, Crithidia) where numerous additions or deletions of uridine residues created tra ...

Complexes composed of RNA and protein play essential roles at multiple levels in the gene expression pathway, including transcription, RNA processing, transport, and translation. In this chapter, we describe a generally applicable system that allows the function of specific RNA-pr ...

The use of in vitro model systems has been essential to the dissection of the mechanisms for transcription regulation. It is now possible to reconstitute specific transcription by each of the three eukaryotic RNA polymerases using purified or at least partially purified protein factors (1 ...

A fundamental goal of gene analysis is to determine what factors regulate the temporal and spatial expression of the gene of interest. This type of analysis requires the identification of the promoter region of the gene, characterization of the specific DNA sequences that regulate the expre ...

Transcriptional control is mediated through interactions involving specific nuclear proteins and target DNA sequences found in the promoters and other regulatory regions of eukaryotic genes. Consequently, the ability to define the location and characterize the nature of these ...

The TATA-binding protein TBP plays a central role in eukaryotic transcription initiation (1, 2). TBP binds to the promoter consensus sequence TATAAA, located approx 30 nucleotides upstream of the transcriptional start site. Once bound, TBP facilitates the assembly of RNA polymerase II a ...

Promoter-specific in vitro transcription by RNA polymerase II is now achievable with cell extracts prepared from a variety of animal and plant sources (1–3). For extracts prepared from mammalian cells, HeLa cells provide many advantages. Large quantities of homogeneous cells can be grown ...

The advent of efficient procedures for site-directed and random in vitro mutagenesis has led to detailed exploration of protein structure-function relationships. Using these approaches, amino acid residues critical for protein activity can be determined in the absence of X-ray cr ...