PMID- 26709593
OWN - NLM
STAT- MEDLINE
DCOM- 20161021
LR  - 20181202
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 106
DP  - 2015 Dec 16
TI  - A Fast Air-dry Dropping Chromosome Preparation Method Suitable for FISH in 
      Plants.
PG  - e53470
LID - 10.3791/53470 [doi]
LID - 53470
AB  - Preparation of chromosome spreads is a prerequisite for the successful 
      performance of fluorescence in situ hybridization (FISH). Preparation of high 
      quality plant chromosome spreads is challenging due to the rigid cell wall. One 
      of the approved methods for the preparation of plant chromosomes is a so-called 
      drop preparation, also known as drop-spreading or air-drying technique. Here, we 
      present a protocol for the fast preparation of mitotic chromosome spreads 
      suitable for the FISH detection of single and high copy DNA probes. This method 
      is an improved variant of the air-dry drop method performed under a relative 
      humidity of 50%-55%. This protocol comprises a reduced number of washing steps 
      making its application easy, efficient and reproducible. Obvious benefits of this 
      approach are well-spread, undamaged and numerous metaphase chromosomes serving as 
      a perfect prerequisite for successful FISH analysis. Using this protocol we 
      obtained high-quality chromosome spreads and reproducible FISH results for 
      Hordeum vulgare, H. bulbosum, H. marinum, H. murinum, H. pubiflorum and Secale 
      cereale.
FAU - Aliyeva-Schnorr, Lala
AU  - Aliyeva-Schnorr L
AD  - Leibniz Institute of Plant Genetics and Crop Plant Research (IPK).
FAU - Ma, Lu
AU  - Ma L
AD  - Leibniz Institute of Plant Genetics and Crop Plant Research (IPK); School of 
      Biological and Chemical Sciences, Queen Mary University of London.
FAU - Houben, Andreas
AU  - Houben A
AD  - Leibniz Institute of Plant Genetics and Crop Plant Research (IPK); 
      houben@ipk-gatersleben.de.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20151216
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (DNA Probes)
SB  - IM
MH  - *Chromosomes, Plant
MH  - DNA Probes/chemistry/genetics
MH  - Hordeum/chemistry/*genetics
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Secale/chemistry/*genetics
PMC - PMC4694022
EDAT- 2015/12/29 06:00
MHDA- 2016/10/22 06:00
PMCR- 2017/12/16
CRDT- 2015/12/29 06:00
PHST- 2015/12/29 06:00 [entrez]
PHST- 2015/12/29 06:00 [pubmed]
PHST- 2016/10/22 06:00 [medline]
PHST- 2017/12/16 00:00 [pmc-release]
AID - 53470 [pii]
AID - 10.3791/53470 [doi]
PST - epublish
SO  - J Vis Exp. 2015 Dec 16;(106):e53470. doi: 10.3791/53470.