遗传学实验技术

Genome mismatch scanning (GMS) is a rapid method of isolating regions of identity by descent (IBD) between two related individuals (1–5). With the availability of simple PCR techniques, vast numbers of highly informative genomewide polymorphic markers, and more recently, radiation hy ...

Comparative genomic hybridization (CGH) provides genome-scale overviews of chromosomal copy number changes in tumors (1). Unlike conventional cytogenetic analysis, it needs no cell culturing, making it applicable to practically any kind of clinical specimen from which DNA can be o ...

For accurate assembly of a bacterial clone map (or contig), it is necessary to precisely detect the degree of overlap and map order of clones. One way is to generate fingerprints for all the clones in a given project. The overlap between clones is determined by the proportion of comigrating restriction ...

Bacterial artificial chromosomes (BACs) represent a very useful cloning system for large DNA fragments and utilize the Escherichia coli F factor as their backbone. E. coli F factor allows strict copy number control of the clones so that they are stably maintained at one to two copies per cell (1). The s ...

The full integration of the cytogenetic, genetic, and physical maps together with the search to identify all the genes of an organism and the effort to position them on the corresponding integrated map, has long been a key issue in genetics. In all three fields of mapping, enormous progress has been made ...

Fluorescence in situ hybridization (FISH) provides a rapid means of placing labeled DNA segments into a wider genomic context. Mapping to banded metaphase chromosomes anchors clones for specific genes or markers in a well-recognized framework and provides a useful confirmation that ...

Massively parallel sequencing technology opens new possibilities for epigenetic research. Many methods have been developed based on the new sequencing platforms, allowing an ultra-deep mapping of epigenetic variants in a fast and cost-effective way. However, handling millions ...

Lysine acetylation of histones is one of the major epigenetic regulators of chromatin conformation and gene expression. The dynamic nature of histone acetylation is determined by the counterbalancing activity of histone acetyltransferase and histone deacetylase (HDAC) enzy ...

DNA methylation, which most commonly occurs at the C5 position of cytosines within CpG dinucleotides, plays a pivotal role in many biological procedures such as gene expression, embryonic development, cellular proliferation, differentiation, and chromosome stability. Aberr ...

The miRBase Sequence database is the primary repository for published microRNA (miRNA) sequence and annotation data. miRBase provides a user-friendly web interface for miRNA data, allowing the user to search using key words or sequences, trace links to the primary literature referenc ...

MicroRNAs (miRNAs) have recently emerged as important regulators of gene expression controlling central biological processes. These small, approx 22-nucleotide (nt)-long RNA molecules induce translational suppression when they are imperfectly matched to their target me ...

Small interefence RNA (siRNA) has revamped the technology of gene silencing in cultured mammalian cells after its first demonstration by Tushl et al. 4 yr ago. To circumvent the cost and the inconvenience in identifying a unique siRNA duplex that can quench target gene expression, we devised a rep ...

MicroRNAs (miRNAs) have a vital role in the generation of plant forms through posttranscriptional regulation of the accumulation of developmental regulators. Analysis of their roles requires detailed knowledge of their expression patterns. We describe an in situ hybridization ...

MicroRNAs (miRNAs) are ubiquitous regulators of gene expression in plants and ani-mals. Their distinctive structure, as very short RNAs with a 5′-phosphate and 3′-hydroxyl group, has enabled the development of protocols to clone miRNAs. After enrichment of these small molecules by size, s ...

MicroRNAs (miRNAs), an abundant class of approx 22-nucleotide (nt) small RNAs that control gene expression at the posttranscriptional level, may play important roles during normal hematopoiesis and leukemogenesis. This chapter focuses on the methods and strategies for dissecti ...

Animal genomes contain on the order of at least hundreds of microRNAs (miRNAs). Although most remain uncharacterized, it is already clear that miRNAs regulate many bio-logical processes. A number of Drosophila miRNAs have been identified as likely cell death regulators, but functions for ...

Initially reported for Caenorhabditis elegans, microRNA (miRNA) has been shown to regulate gene expression in plants, flies, and mammals (1,2). Recently, findings of miRNA have been extended to viruses (3,4). Here, we report on our approaches to investigate the role of miRNAs in human immunode ...

MicroRNAs (miRNAs) are 21- to 25-nucleotides (nt) long and interact with messenger RNAs to trigger either translational repression or RNA cleavage through RNA interference (RNAi). We have shown that HIV-1 nef double-stranded RNA from AIDS patients who are long-term nonprogressors, inh ...

Fragile X syndrome, a common form of inherited mental retardation, is caused by the loss of the Fragile X mental retardation protein (FMRP). FMRP, which may regulate translation in neurons, not only associates with specific messenger RNAs (mRNAs) and with microRNAs (miRNAs), but also associa ...

Eukaryotic CDC6 gene function is required for the initiation of DNA replication and is a key regulatory protein during cell cycle progression. The human CDC6 gene is not expressed in most normal tissues, in contrast with its marked expression in proliferating cancer cells. An effective way to e ...