PMID- 20809329
OWN - NLM
STAT- MEDLINE
DCOM- 20101214
LR  - 20131121
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 659
DP  - 2010
TI  - Fluorescence in situ hybridization with Bacterial Artificial Chromosomes (BACs) 
      to mitotic heterochromatin of Drosophila.
PG  - 389-400
LID - 10.1007/978-1-60761-789-1_30 [doi]
AB  - The organization of eukaryotic chromosomes into euchromatin and heterochromatin 
      represents an enigmatic aspect of genome evolution. Constitutive heterochromatin 
      is a basic, yet still poorly understood component of eukaryotic genomes and its 
      molecular characterization by means of standard genomic approaches is 
      intrinsically difficult. Drosophila melanogaster polytene chromosomes do not seem 
      to be particularly useful to map heterochromatin sequences because the typical 
      features of heterochromatin, organized as it is into a chromocenter, limit 
      cytogenetic analysis. In contrast, constitutive heterochromatin has been 
      well-defined at the cytological level in mitotic chromosomes of neuroblasts and 
      has been subdivided into several bands with differential staining properties. 
      Fluorescence in situ hybridization (FISH) using Bacterial Artificial Chromosomes 
      (BAC) probes that carry large genomic portions defined by sequence annotation has 
      yielded a "revolution" in the field of cytogenetics because it has allowed the 
      mapping of multiple genes at once, thus rendering constitutive heterochromatin 
      amenable to easy and fast cytogenetics analyses. Indeed, BAC-based FISH 
      approaches on Drosophila mitotic chromosomes have made it possible to correlate 
      genomic sequences to their cytogenetic location, aiming to build an integrated 
      map of the pericentric heterochromatin. This chapter presents our standard 
      protocols for BAC-based FISH, aimed at mapping large chromosomal regions of 
      mitotic heterochromatin in Drosophila melanogaster.
FAU - Accardo, Maria Carmela
AU  - Accardo MC
AD  - Dipartimento di Genetica e Biologia Molecolare and Istituto Pasteur-Fondazione 
      Cenci-Bolognetti, Universita "La Sapienza", Rome, Italy.
FAU - Dimitri, Patrizio
AU  - Dimitri P
LA  - eng
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (DNA, Bacterial)
RN  - 0 (Heterochromatin)
RN  - 0 (Rhodamines)
RN  - NQ1SX9LNAU (Digoxigenin)
SB  - IM
MH  - Animals
MH  - Brain/cytology/metabolism
MH  - Chromosomes, Artificial, Bacterial/*metabolism
MH  - Chromosomes, Insect/metabolism
MH  - DNA, Bacterial/isolation & purification/metabolism
MH  - Digoxigenin/metabolism
MH  - Drosophila melanogaster/*cytology/*metabolism
MH  - Heterochromatin/*metabolism
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Larva
MH  - *Mitosis
MH  - Rhodamines/metabolism
EDAT- 2010/09/03 06:00
MHDA- 2010/12/16 06:00
CRDT- 2010/09/03 06:00
PHST- 2010/09/03 06:00 [entrez]
PHST- 2010/09/03 06:00 [pubmed]
PHST- 2010/12/16 06:00 [medline]
AID - 10.1007/978-1-60761-789-1_30 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2010;659:389-400. doi: 10.1007/978-1-60761-789-1_30.