PMID- 11269343
OWN - NLM
STAT- MEDLINE
DCOM- 20010517
LR  - 20190915
IS  - 0831-2796 (Print)
IS  - 0831-2796 (Linking)
VI  - 44
IP  - 1
DP  - 2001 Feb
TI  - An efficient method for the physical mapping of transgenes in barley using in 
      situ hybridization.
PG  - 104-10
AB  - The genetic transformation of crops by particle bombardment and Agrobacterium 
      tumefaciens systems have the potential to complement conventional plant breeding 
      programmes. However, before deployment, transgenic plants need to be 
      characterized in detail, and physical mapping is an integral part of this 
      process. Therefore, it is important to have a highly efficient method for 
      transgene detection by fluorescence in situ hybridization (FISH). This study 
      describes a new approach, which provides efficient control of probe length and 
      labelling, both of which play an important role in in situ hybridization of 
      transgenes. The approach is based on reducing the size of the plasmid prior to 
      labelling by nick translation, rather than using the whole or linearized plasmid, 
      or varying the amounts of DNaseI in the nick translation mixture. This provided 
      much more efficient labelling of the probe, which yielded optimal hybridization. 
      minimal fluorescent background, and accurate physical location of the transgene.
FAU - Salvo-Garrido, H
AU  - Salvo-Garrido H
AD  - John Innes Centre, Norwich Research Park, UK.
FAU - Travella, S
AU  - Travella S
FAU - Schwarzacher, T
AU  - Schwarzacher T
FAU - Harwood, W A
AU  - Harwood WA
FAU - Snape, J W
AU  - Snape JW
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Canada
TA  - Genome
JT  - Genome
JID - 8704544
RN  - 0 (DNA Probes)
SB  - IM
MH  - DNA Probes
MH  - *Genes, Plant
MH  - Hordeum/*genetics
MH  - In Situ Hybridization/*methods
MH  - Physical Chromosome Mapping/*methods
MH  - *Transgenes
EDAT- 2001/03/28 10:00
MHDA- 2001/05/18 10:01
CRDT- 2001/03/28 10:00
PHST- 2001/03/28 10:00 [pubmed]
PHST- 2001/05/18 10:01 [medline]
PHST- 2001/03/28 10:00 [entrez]
AID - 10.1139/gen-44-1-104 [doi]
PST - ppublish
SO  - Genome. 2001 Feb;44(1):104-10. doi: 10.1139/gen-44-1-104.