Change search
ReferencesLink to record
Permanent link

Direct link
The CIC YAC library: Sizing of the clones and determination of clones carrying repeated DNA sequences.
Show others and affiliations
1995 (English)In: The Plant Journal, ISSN 0960-7412, E-ISSN 1365-313X, Vol. 8, no 5, 763-770 p.Article in journal (Refereed) PublishedText
Abstract [en]

A new Arabidopsis thaliana (ecotype Columbia) genomic library has been constructed in Yeast Artificial Chromosomes: the CIC library (for CEPH, INRA and CNRS). Optimization of plant culture conditions and protoplast preparation allowed the recovery of large amounts of viable protoplasts. Mechanical shearing of DNA was minimized by isolation of DNA from protoplasts embedded in agarose. Cloning of large inserts was favored by including two successive size fractionation steps (after partial EcoRI digestion and after ligation with the vector arms), which selected DNA fragments larger than 350 kb. The library consists of 1152 clones with an average insert size of 420 kb. Clones carrying chloroplast DNA and various nuclear repeated sequences have been identified. Twenty-one per cent of the clones are found to contain chloroplast DNA. Therefore, the library represents around four nuclear genome equivalents. The clones containing 5S rDNA genes, 18S-25S rDNA sequences and the 180 bp paracentromeric repeated element account for 3.6%, 8.9% and 5.8%, respectively. Only one clone was found to carry the 160 bp paracentromeric repeated element. Given the smaller size of clones carrying Arabidopsis repeated DNA, the average size of remaining clones is around 480 kb. The library was screened by PCR amplification using pairs of primers corresponding to sequences dispersed in the genome. Seventy out of 76 pairs of primers identified from one to seven YAC clones. Thus at least 92% of the genome is represented in the CIC library. The survey of the library for clones containing unlinked DNA sequences indicates that the proportion of chimeric clones is lower than 10%.

Place, publisher, year, edition, pages
1995. Vol. 8, no 5, 763-770 p.
National Category
Natural Sciences
URN: urn:nbn:se:umu:diva-114194DOI: 10.1046/j.1365-313X.1995.08050763.xOAI: diva2:914097

Times Cited: 146 147

Available from: 2016-03-23 Created: 2016-01-15 Last updated: 2016-03-23

Open Access in DiVA

No full text

Other links

Publisher's full texthttp://<Go to ISI>://WOS:A1995TF90800014

Search in DiVA

By author/editor
Bellini, C.
In the same journal
The Plant Journal
Natural Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 12 hits
ReferencesLink to record
Permanent link

Direct link