PMID- 21326371
OWN - NLM
STAT- MEDLINE
DCOM- 20110609
LR  - 20161020
IS  - 1480-3321 (Electronic)
IS  - 0831-2796 (Linking)
VI  - 54
IP  - 2
DP  - 2011 Feb
TI  - High-density fluorescence in situ hybridization signal detection on barley 
      (Hordeum vulgare L.) chromosomes with improved probe screening and reprobing 
      procedures.
PG  - 151-9
LID - 10.1139/G10-098 [doi]
AB  - The barley (Hordeum vulgare L.) genome was screened to identify sequences that 
      could be used for fluorescence in situ hybridization (FISH). From 2000 
      transformed bacterium colonies carrying barley clones, 56 colonies were selected 
      on the basis of the patterns that their PCR products produced when subjected to 
      agarose gel electrophoresis. Among them, 42 (75%) exhibited fluorescent signals 
      on barley chromosomes after in situ hybridization using the directly labeled PCR 
      products. Sequencing revealed seven clones, pHv-365, pHv-177, pHv-1112, pHv-689, 
      pHv-1476, pHv-1889, and pHv-1972, to be newly identified FISH-positive sequences. 
      The remainder possess previously described sequences such as 5S, GAA 
      microsatellite, centromere repeats, HVT01, and pHvMWG2315 (324 bp repeat). It is 
      shown here that a combination of five probes, which produce strong signals on 
      barley chromosomes, pHv-38 (5S), pHv-365, pHv-961 (HVT01), GAA, and TAG 
      microsatellites, offer unequivocal recognition of each chromosome. The 
      combination of three probes, i.e., pHv-1123 (barley 324 bp repeat), GAA, and TAG, 
      decorated entire chromosomes with fine dotted signals and was useful for 
      detecting the break points of aberrant chromosomes. The signals' distributions of 
      pHv-177, pHv-1112, and TAG were highly polymorphic. An improved reprobing 
      procedure and its usefulness are also discussed.
FAU - Kato, Akio
AU  - Kato A
AD  - Laboratory of Plant Breeding, Faculty of Life and Environmental Sciences, Kyoto 
      Prefectural University, 1-5 Shimogamo Hangi-cho, Sakyo-ku, Kyoto-shi, Kyoto 
      606-0823, Japan. katoa@kpu.ac.jp
LA  - eng
PT  - Journal Article
PL  - Canada
TA  - Genome
JT  - Genome
JID - 8704544
RN  - EC 3.1.- (Deoxyribonucleases)
SB  - IM
MH  - Centromere/genetics
MH  - Chromosomes, Plant/*genetics
MH  - Deoxyribonucleases/metabolism
MH  - Escherichia coli/cytology
MH  - *Genome, Plant
MH  - Hordeum/*genetics
MH  - *Hybridization, Genetic
MH  - Microsatellite Repeats
MH  - Sequence Analysis, DNA/*methods
EDAT- 2011/02/18 06:00
MHDA- 2011/06/10 06:00
CRDT- 2011/02/18 06:00
PHST- 2011/02/18 06:00 [entrez]
PHST- 2011/02/18 06:00 [pubmed]
PHST- 2011/06/10 06:00 [medline]
AID - g10-098 [pii]
AID - 10.1139/G10-098 [doi]
PST - ppublish
SO  - Genome. 2011 Feb;54(2):151-9. doi: 10.1139/G10-098.