Analysis and sorting of chromosomes by flow cytometry: New trends

Metezeau, Philippe; Schmitz, Annette; Frelat, G.

doi:10.1016/0248-4900(93)90112-r

Cited by 11 publications

(8 citation statements)

References 71 publications

(124 reference statements)

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“…This has been achieved by¯ow sorting (Metezeau et al 1993;Arumaganathan et al 1994;Dolezel et al 1994;Ellegren and Basu 1995;Veuskens et al 1995;Dolezel and Lucretti 1995) or chromosome dissection (Busch et al 1995;Chen and Armstrong 1995). Both of these techniques have signi®cant limitations.…”

Section: Discussionmentioning

confidence: 99%

“…In plants the main techniques available for the construction of chromosome-speci®c clone libraries have been chromosome micro-dissection (Busch et al 1995;Chen and Armstrong 1995) or¯ow cytometry (reviewed by Dolezel et al 1994 andMetezeau et al 1993). These techniques have allowed the construction of chromosome-and chromosome arm-speci®c libraries (Arumuganathan et al 1994;Chen and Armstrong 1995;Veusekens et al 1995), although in some cases only enriched libraries could be generated.…”

Section: Introductionmentioning

confidence: 99%

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Isolation of lambda and YAC clones from defined regions of the rye genome

Langridge

Guo²,

Francki³

1998

Mol Gen Genet

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A dispersed, rye-specific element has been used to isolate clones of rye origin from wheat plants containing only a single rye chromosome arm or segment. In this way a set of 23 YAC clones has been isolated from the short arm of rye chromosome 1 (1RS). This technique was extended to isolate clones from a small region of 1RS that contains a large number of agronomically important genes. The targeted cloning method allowed the isolation of 26 classes of lambda clones representing about 5% of the region. Ten of the lambda clones could be mapped to segments within this region. A third example of the application of this technique involved the isolation of clones from a very small but fully functional rye chromosome, the midget chromosome. These clones have allowed the confirmation of the origin of the midget from 1RL, and may provide a tool for the isolation of structural elements of cereal chromosomes. This technique allows the identification of clone libraries for any rye chromosome or chromosome arm, since substitution, addition and translocation lines are available for all rye chromosomes. Furthermore, the technique allows isolation of clones derived from segments of the rye genome recombined into wheat. The method is technically simple and both lambda and YAC libraries can be constructed. Synteny between the genomes of the cereals allows region-specific libraries from rye to be used to target regions of the wheat and barley genomes.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Isolation of lambda and YAC clones from defined regions of the rye genome

Langridge

Guo²,

Francki³

1998

Mol Gen Genet

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“…In analysis of small bacteria, careful attention should be paid to optimisation of instrumental set-up and to the quality of sheath and microbial suspension buffers. As for chromosome analysis, great care must be taken in optical alignment, choice of filters and optical cleanness [75,122]. Good instruments with built-in logarithmic amplifiers are a must.…”

Section: Algaementioning

confidence: 99%

Recent advances of flow cytometry in fundamental and applied microbiology

Fouchet

Jayat

Héchard³

et al. 1993

Biology of the Cell

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This review focuses on the recent applications of flow cytometry (FCM) in microbiological research (1987-mid 1992). It tries to give a scope of the important breakthroughs which occurred in this field during this period. The technical difficulties of microorganism analysis by flow cytometry is briefly appraised. The significance and the limits of the different microbial cell parameters attainable by flow analyses are systematically evaluated: light scatter for cell size and structure, fluorescence measurements for quantification of cellular components, microbial antigen detection and cell physiological activity estimation. Emphasis is given on the new technological advances which appeared in the last two years. The second part of the review is devoted to the analysis of the usefulness of flow cytometric approach in the different fields of microbiology: fundamental studies in microbial physiology, differentiation, microbial ecology and aquatic sciences, medical microbiology, parasitology, microbial pharmacology and biotechnology.

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“…Flowcytometry, while a valuable method for isolating some of the large chromosomes, is limited in the preparation of suspensions for intact chromosomes, in sensitivity for isolating small chromosomes, and in sorting single chromosomes (Doležel et al, 1994;Lee et al, 1997;Métézeau, et al, 1993). Chromosome microdissection and microcloning are valuable techniques for dissecting target chromosome or chromosome segments and for cloning the dissected chromosomes in order to construct chromosome specific libraries.…”

Section: Introductionmentioning

confidence: 99%

Development of microdissection and chromosome specific genomic library in Lilium tigrinum

Hwang

Lim

2011

Genes Genom

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Techniques for microdissection and microcloning were established using chromosome 1 of triploid Lilium tigrinum. Chromosome 1 was dissected from a membrane slide using a microbeam system. Digestion with proteinase K was done before PCR amplification for more than 24 hours. The dissected chromosomes were then amplified by degenerate oligonucleotide primed PCR (DOP-PCR) and linker adaptor-mediated PCR (LA-PCR). Successful PCR amplification relied on critical concentrations of both MgCl2 and Taq polymerase. The optimum concentration of MgCl2 and Taq polymerase was 2.5 mM and 0.5 U, respectively. Amplification of the dissected chromosome using DOP-PCR had more sensitivity dependent upon PCR factors, but LA-PCR was more dependent on the linker ligation. Amplified DNA products ranged from 100 to 2500 bp both for DOP-and LA-PCR. Evaluated clones only ranged from 100 to 1700 bp for DOP-PCR and 100 to 900 bp for LA-PCR. Based on the sequence results, most of the sequences from the DOP-PCR and LA-PCR showed no significant similarity with known data in NCBI database. However, about 2% of the sequence data was partially matched with plant microsatellites with low similarity. The results derived from the microdissection of a large genome organism such as Lilium showed informative and useful for the development of microsatellite repeat markers. Sequence data from the chromosome specific DNA library was considered for the development of microsatellite markers.

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Analysis and sorting of chromosomes by flow cytometry: New trends

Cited by 11 publications

References 71 publications

Isolation of lambda and YAC clones from defined regions of the rye genome

Isolation of lambda and YAC clones from defined regions of the rye genome

Recent advances of flow cytometry in fundamental and applied microbiology

Development of microdissection and chromosome specific genomic library in Lilium tigrinum

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