Genomic imprinting is a regulatory mechanism by which, at certain gene loci, one of the two alleles becomes repressed according to its parental origin. This chapter focuses on mammalian imprinting, which regulates not only the parental allele-specific gene expression on autosomal chr ...
A functional centromere is formed by a chromosomal domain that very often, but not always, is recognizable by a primary constriction in metaphasic chromosomes. It is associated with a kinetochore through which a link is established with the microtubules, which pull the sister chromatids to ...
DNA replication is the key process for the conservation and transmission of genetic information in all living organisms. The bi-directional DNA replication does not start randomly along the genome but rather at precise chromosomal regions, defined as origins of DNA replication where, a ...
Barring exceptional instances, the DNA contained in eukaryotic chromosomes is linear. Linearity of the chromosomal DNA and the compartmentalized architecture of the eukaryotic cell are the two principle features that distinguish the prokaryotes from the eukaryotes and that have ...
Telomerization is an operational term for appparent immortalization sustained by prevention of telomere shortening. It is always necessary to use the term apparent because immortalization, if taken literally, requires one to demonstrate that cells divide forever, a property that ...
The functional organization of the eukaryotic chromosome was first elucidated at a molecular level in the budding yeast, Saccharomyces cerevisiae, providing the basis for the successful creation of yeast artificial chromosomes (YACs) (1). The structures that confer chromosome f ...
Subtractive cloning is an ideal technique for identifying genes differentially expressed in two nuclear acids population (1). The polymerase chain reaction (PCR)-based subtraction is the method of choice when the starting samples are heterogeneous or difficult to obtain, which of ...
Heat will separate or "e;melt"e; double-stranded DNA into single-stranded DNA by disrupting its hydrogen bonds. T m (melting temperature) is the temperature at which half the DNA strands are single-stranded and half are double-stranded. T m characterizes the stability of the DNA h ...
Polymerase chain reaction (PCR) optimization and troubleshooting can consume considerable energy and resources because of the finicky and often unpredictable nature of the reactions. Small variations in any of the many variables in a given reaction can have a pronounced effect on the re ...
Long polymerase chain reaction (PCR)(1 5), specifically, XL PCR (Extra-Long Polymerase Chain Reaction), has enabled amplification of expanded trinucleotide repeats of the neuromuscular disease myotonic dystrophy (6) a 9-kb HIV-1 provirus from primary isolate DNA (7), 24.2-kb fragm ...
Although Thermus aquaticus (Taq) and Thermus thermophilus (Tth) DNA polymerases have the ability to reverse transcribe RNA to complementary DNA (cDNA) and subsequently amplify the target cDNA, they are not usually the first choices for reverse transcription-polymerase chain rea ...
Many common molecular biology techniques including polymerase chain reaction (PCR) (1), Southern blotting (2), comparative genomic hybridization (3), and in situ hybridization (4) have been adapted for use with paraffin-embedded tissue (PET). PCR-amplified products from PET can be ...
Polymerase chain reaction (PCR) has been applied to the amplification of long DNA fragments from a variety of sources, including genomic, mitochondrial, and viral DNAs (1-5). We have adapted the concept of long PCR technology to reverse-transcription (RT) PCR (6). Here, we describe the paramet ...
The amplification of GC-rich templates by any PCR method is usually a difficult task and despite the development of modified methods and conditions, this type of amplification still remains a specific case approach. Problems usually observed with GC-rich DNA are constraint of template am ...
A successful cloning of polymerase chain reaction (PCR)-derived DNA fragment is a key step for further analysis of the amplified DNAs, though it is often a difficult task. Many cloning methods have been established; various commercial cloning kits are also available. These methods can be sepa ...
Microsatellites, also referred to as short tandem repeats (STR) or simple sequence repeats (SSR), are highly polymorphic and abundant sequences dispersed throughout most eukaryotic nuclear genomes (1-3). In recent years, microsatellites have been used for linkage map constructi ...
Numerous techniques have been developed for the cloning of polymerase chain reaction (PCR) products. These include the incorporation of restriction enzyme sites into the PCR primers (1), blunt-end cloning (2,3), TA cloning (4,5), ligation-independent cloning (LIC) (6-11), and in vivo clo ...
The polymerase chain reaction (PCR) technique has proved to be a powerful tool for rapid amplification of DNA fragments of interest during cloning. Insertion of PCR products into suitable vectors in order to construct plasmids for protein expression, or to create chimeric genes or to study in ...
The expression of cloned genes in prokaryotic or eukaryotic host cells provides the means not only for the study of gene function but also for the production of substantial amounts of protein and nonprotein molecules for commercial and investigational use. In the case of proteins, strategies ...
Efficient strategies for the production of recombinant proteins are gaining increasing importance, as more applications that require high amounts of high-quality proteins reach the market. Higher production efficiencies and, consequently, lower costs of the final product are ...
