细胞功能测定

Genomic amplification with transcript sequencing (GAWTS) (1,2) is a generally applicable method for direct sequencing of PCR material. GAWTS is centered around the attachment of a phage promoter sequence (T7, Sp6, or T3) to the 5′-end of one or both PCR primers. The phage promoter sequence allows t ...

Because only between 1 and 10% of bacteria present in soil and aquatic environments are culturable using currently available methods, attempts to identify and quantify the nonculturable, majority population must circumvent the need for culture. Molecular biological techniques, ...

The advent of direct sequencing of polymerase chain reaction (PCR) products has permitted extremely rapid analysis of DNA mutants and cDNA clones. However, direct PCR sequencing has been problematic for a number of technical reasons, including the presence of impurities and excess olig ...

The eukaryotic cell cycle is a process in which cells grow and then divide into two genetically identical cells. The cell cycle is divided into four discrete phases allowing for the orderly transition from deoxyribonucleic acid (DNA) replication to chromosomal condensation, spindle fo ...

Haploid yeast cells mate to form a zygote, whose progeny are diploid cells. A fundamentally sexual event, related to fertilization, yeast mating nevertheless exhibits cytological properties that appear similar to somatic cell fusion. A large collection of mutations that lead to defec ...

Hyphal fusion occurs at different stages in the vegetative and sexual life cycle of filamentous fungi. Similar to cell fusion in other organisms, the process of hyphal fusion requires cell recognition, adhesion, and membrane merger. Analysis of the hyphal fusion process in the model organi ...

Differentiation of vegetative cells of the haploid eukaryote Chlamydomonas is dependent on environmental conditions. Upon depletion of nitrogen and exposure to light, vegetative cells undergo a mitotic division, generating gametes that are either mating-type plus (mt) or mati ...

In the nematode Caenorhabditis elegans, 300 of the 959 somatic nuclei present in the adult hermaphrodite are located in syncytia. These syncytia are formed by the fusion of mononucle-ate cells throughout embryonic and postembryonic development. These cell fusions occur in a well-char ...

Myogenic differentiation in Drosophila melanogaster, as in many other organisms, involves the generation of multinucleate muscle fibers through the fusion of myoblasts. Prior to fusion, the myoblasts become specified as one of two distinct cell types. They then become competent to fu ...

Direct sequencing of PCR products (1) has proven to be a powerful method in the generation of nucleic acid sequence data. Using these techniques, it is possible to produce microgram quantities of pure target DNA and subsequently its nucleotide sequence in a few hours, even, theoretically, from one ...

Following the isolation of a clonal recombinant phage or plasmid after screening a cDNA library, the first analyses that are routinely performed are the determination of the insert size and the sequence of the 3′- and 5′-ends of the cloned fragment. Prior to the use of PCR, this was a laborious process, esp ...

The following protocol describes a method that can be used for the direct sequencing of polymerase chain reaction (PCR) products using nonradioactive detection procedures. It is based on our experience with a method we have previously published (1). Direct sequencing of PCR products is gen ...

The use of magnetic particles in many fields of biochemistry, molecular biology, and medicine has been well documented, and several magnetic particles are now available for diagnostic and cell-separation purposes. The solid-phase approaches have improved robustness by their incr ...

The time and labor required for generating nucleotide sequence information has been significantly reduced since the development of the polymerase chain reaction (PCR). PCR allows the production of sufficient amounts of DNA template in vitro, and consequently, the time-consuming c ...

Understanding cell growth and cell division involves the study of regulatory events that occur in a cell cycle phase-dependent manner. Studies analyzing cell cycle regulatory mechanisms and cell cycle progression invariably require synchronization of cell populations at spec ...

Synchronized populations of large numbers of cells can be obtained by centrifugal elutriation on the basis of sedimentation properties of small round particles, with minimal perturbation of cellular functions. The physical characteristics of cell size and sedimentation veloc ...

Widespread interest in cell synchronization is maintained by the studies of control mechanisms involved in cell cycle regulation. During the synchronization distinct subpopulations of cells are obtained representing different stages of the cell cycle. These subpopulations ...

The cell cycle is controlled by complex regulatory network to ensure that the phases of the cell cycle happen in the right order and transitions between phases happen only if the earlier phase is properly finished. This regulatory network receives signals from the environment, monitors the st ...

Septic syndromes represent a major, although largely under-recognized, healthcare problem worldwide accounting for thousands of deaths every year. Although flow cytometry (FCM) remains a relatively confidential diagnostic tool, it is useful at every step of intensive care unit ( ...

The efficient use of haematopoietic stem cells (HSC) for transplantation is often limited by the relatively low numbers of HSC collected. The ex vivo expansion of HSC for clinical use is a potentially valuable and safe approach to increase HSC numbers thereby increasing engraftment and redu ...