The technique of chromosome microdissection and microcloning has been developed for more than 20 years. As a bridge between cytogenetics and molecular. Microdissection of the A h01 chromosome in upland cotton and microcloning of resistance gene anologs from the single chromosome. The aim of this paper is to review the different techniques available for chromosome microdissection and microcloning, and their use for the.
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Chromosome microdissection – Wikipedia
The DOP-PCR technique, which is rapid, efficient, and species-independent, is designed to amplify target DNAs at frequently occurring priming sites using the primer of partially degenerate sequence, without restrictions imposed by the complexity or the origin of DNA.
Direct chromosomal microdissection and microcloning is a rapid technique for providing probes for such areas.
Finally, a two-step PCR amplification is performed to select the hybridized fraction of the samples: However, the present state of microdissection and microcloning technique cannot meet the needs for sequencing the complete specific chromosome DNA due to low coverage and small size of inserted DNA fragments.
Zuo Wu Xue Bao. A novel method to isolate the common fraction of two DNA samples: Or, a portion of a chromosome may be repeated over and over again, resulting in an unusually wide, dark band known as a homogeneously staining region.
Molecular cloning of DNA from specific chromosomal regions by microdissection and sequence-independent amplification of DNA. Analysis and sorting of chromosmes by flow cytometry: Studies on contamination of cytoplasm DNA and its control in plant chromosome microdissection.
The purpose of this article is to review the development of this technique and its application in the field of genomic research. To construct a high specificity, high coverage and low bias library with larger insert fragments, we only need single chromosome DNA. In order to isolate one special chromosome, Griffin et al. The earliest chromosome microcloning [ 61 ] was a kind of direct cloning of the dissected chromosomal material in a nanoliter microdrop contained in an oil chamber.
However, the introduction of PCR technology to the micro-cloning protocols has brought a substantial improvement in cloning efficiency [ 46 ]. During those 20 years, this technique has not only been benefited from other technological advances but also cross-fertilized with other techniques.
Views Read Edit View history. It is a new potential approach developed by our lab to rapidly generate ESTs from a specific chromosome or even a chromosome-specific-region paper in preparation.
Scientists who study chromosomes are known as cytogeneticists.
This is a good method for the enrichment of ESTs of the specific chromosomes or chromosome fragments [ 1939 ]. The probes usually generate from the clones of cosmid library, yeast artificial chromosome YAC and bacterial artificial chromosome BAC.
Journal List Curr Genomics v. Micro-and macrochromosome micrrodissection generated by flow cytometry mircocloning microdissection: Refined examination of plant metaphase chromosome structure at different levels made feasible by new isolation methods. Today, it becomes microclohing practicality with extensive uses. Microdissection and PCR amplification of single soybean chromosome.
Acid Treatment Leads to Depurination One of the major concerns for the successful application of chromosome microdissection and microcloning technique was the extensive depurination of the chromosomal DNA caused by acid treatment during the sample fixation in chromosome preparation [ 14 ].
Chen Q, Armstrong K. Fluorescence in situ hybridization FISH plays an essential role in research and clinical diagnostics.
For example, one chromosome may have a piece of another chromosome inserted within it, creating extra bands. Short cuts for genomic walking: A New Method of Ests Selection may be more Efficient — Hybrid Suppressive Amplification Selection It is a new potential approach developed by our lab to rapidly generate ESTs from a specific chromosome or even a chromosome-specific-region paper in preparation.
Sci China, C, Life Sci. Author information Article notes Copyright and License information Disclaimer. One of the major concerns for the successful application chromosomr chromosome microdissection and microcloning technique was the extensive depurination of the chromosomal DNA caused by acid treatment during the sample fixation in chromosome preparation [ 14 ].
This page was last edited on 11 Februaryat However, the micordissection is huge and highly redundant, and it is difficult to identify the chromosome specific ESTs from cDNA libraries. They reported that the single cell E. Targeted generation of 16 sequence-tagged sites for bovine chromosome region 5qq25 by microdissection.
Isolation of the single chromosome with BYDV resistant gene.
Benefited from other Techniques Chromosome microdissection and microcloning has been benefited from technological advances and coupling with other techniques, which further improved its application. Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes.