遗传学实验技术

For routine genetic analysis of Cannabis sativa, two methods are currently in use, (a) AFLP; amplified fragment length polymorphism analysis and (b) STR; short tandem repeat analysis. The AFLP method used on capillary electrophoresis instrumentation is fully described in this chapter. ...

STR analysis of canine-derived biological evidence for the identification of individuals is becoming an important tool for forensic investigations. A protocol for the multiplex PCR amplification and capillary electrophoresis of nine autosomal STRs and two fixed-size markers ...

The interpretation of multilocus STR profiling is a complex task that requires a high degree of expertise due to the high number of variables that can affect a biological and analytical process like this. The purpose of this chapter is to provide guidelines to categorize the mixtures using a discr ...

Low template (LT) DNA testing is a more sensitive method of PCR DNA typing which tests lower quantities of DNA compared to traditional PCR DNA protocols. Methods applied in this testing involve amplification or postamplification efforts to increase detection sensitivity. Establish ...

STR analysis of DNA extracted from skeletal samples can play an important role in the identification of missing persons. Here we present a method for the extraction of DNA from skeletal samples involving complete demineralization and digestion of the sample, followed by purification by si ...

The analysis of cell-specific mRNA expression is a promising new method for the identification of body fluids. A number of mRNA markers have been identified for the forensically most relevant body fluids: blood, saliva, semen, vaginal secretions, and menstrual blood. Apart from a significa ...

The typing of single nucleotide polymorphisms (SNPs) located throughout the human mitochondrial genome assists in resolving individuals with an identical HV1/HV2 haplotype. A set of 11 sites which were selected for distinguishing individuals of a common Western European Caucas ...

This work describes the main advantages and the steps involved in the optimization of a multiplex system able to characterize 38 noncoding biallelic Insertion Deletion Polymorphisms(Indels). With this methodology, all markers are amplified in a single PCR, using short amplicons (up to ...

Advancements in high-throughput technologies to measure increasingly complex biological phenomena at the genomic level are rapidly changing the face of biological research from the single-gene single-protein experimental approach to studying the behavior of a gene in the con ...

BioTapestry is an open source, freely available software tool that has been developed to handle the �challenges of modeling genetic regulatory networks (GRNs). Using BioTapestry, a researcher can �construct a network model and use it to visualize and understand the dynamic behavior of a co ...

In recent years, new techniques have spurred the discovery of cis-regulatory DNA elements. These stretches of noncoding DNA contain combinations of recognition sites to which transcription factors (TFs) bind, and in doing so, these TFs can activate or repress transcription. These prot ...

Transcription factors regulate transcription by binding to regulatory regions of genes including the promoter. Few of the transcription factors are well characterized, and few promoters have been described in detail. New methods have been developed to improve both transcription ...

Gene expression regulation is a fundamental biological process leading to complete organism development by controlling processes like cell type specification and differentiation. The accuracy of this process is �governed by transcription factors (TFs) acting within a compl ...

Binding of transcription factors to promoters is a necessary step to initiate transcription. From an evolutionary standpoint, the regulatory proteins and their binding sites are considered to have molecularly coevolved. We developed an efficient yeast strategy, an “inverse one- ...

Chromatin immunoprecipitation experiments followed by ultra-high-throughput sequencing (ChIP-seq) is becoming the method of choice to identify transcription factor binding sites in prokaryotes and eukaryotes in vivo. Here, we review the computational steps that are neces ...

Chromatin immunoprecipitation (ChIP) is a commonly used technique to detect the in vivo binding of proteins to DNA. ChIP is now routinely paired to microarray analysis (ChIP-chip) or next-generation sequencing (ChIP-Seq) to profile the DNA occupancy of proteins of interest on a genome-wi ...

Accurately assessing the binding of transcription factors to cis-regulatory elements in vivo is an essential step toward understanding the mechanisms that govern embryonic development. Genome-wide transcription factor location analysis has been facilitated by the devel ...

Eukaryotic transcription is tightly regulated by transcriptional regulatory elements, even though these elements may be located far away from their target genes. It is now widely recognized that these regulatory elements can be brought in close proximity through the formation of chr ...

DNaseI-hypersensitive sites within chromatin are indicative of genomic loci with regulatory function. Several techniques have been described for analyzing these regions, but are either laborious, offer low-throughput possibilities, or are expensive. We have developed a new a ...

We provide here a protocol for the preparation of cap-analysis gene expression (CAGE) libraries, which allows for measuring the expression of eukaryotic capped RNAs and simultaneously map the promoter regions. The presented protocol simplifies the previously published ones and mo ...