9‐Aminoacridine: An efficient reagent to improve human and plant chromosome banding patterns and to standardize chromosome image analysis

Abstract

BackgroundSuccessful automated chromosome analysis requires the development of new techniques to increase and standardize chromosome length and improve banding patterns.MethodsHuman and plant cells were pretreated with the DNA intercalator 9‐aminoacridine (9‐AMA), and chromosomes were stained with GTG and aceto‐orcein banding techniques and investigated by an image analysis system.ResultsThe human optimal chromosome spreads with the 850 G‐band resolution level, suitable for image analysis, were obtained by 9‐AMA pretreatment for 1 h at a final concentration of 0.5–1 μg/ml, as compared with 600–700 bands after ethidium bromide treatment and about 400 bands without pretreatment. The best results for plant chromosomes were obtained after pretreatment with 1–2 μg/ml of 9‐AMA for 12–24 h. The chromosomes elongated approximately 1.5‐fold, and the resolution of chromosome banding patterns increased, reaching approximately 140 bands per haploid set in the case of camomile.Conclusions9‐AMA is an efficient reagent for the standardization and increasing the resolution of chromosome banding patterns in human and plant chromosomes. It is extremely important for chromosome investigation in small plants. Cytometry Part A 51A:52–57, 2003. © 2002 Wiley‐Liss, Inc.

Comparison of genomes of eight species of sections Linum and Adenolinum from the genus Linum based on chromosome banding, molecular markers and RAPD analysis

“…Root tips (of 0.5 cm) were excised and treated overnight (16-20 h) in an ice-cold water with 1 lg/ml 9-aminoacridine (Sigma) to harvest elongated chromosomes (Muravenko et al 2003). After pretreatment, root tips were fixed in ethanol:acetic acid (3:1) for 3-24 h at room temperature.…”

Karyotype Study of 5 Species of the Family Viverridae in Thailand by High Resolution GTG-Banding Technique

“…Several chromosome anti-contraction agents are available and include ethidium bromide, the thymidine analogue BrdU, 5-azacytidin, actinomycin D, echinomycin, and others (Lawce andBrown 1997, Sawyer 1995). Recently, 9-aminoacridine was used to increase chromosome spreads to 850 BPHS resolution, compared to 600-700 BPHS with ethidium bromide (Muravenko et al 2003, Lim et al 2004). …”

A new chromosome fluorescence banding technique combining DAPI staining with image analysis in plants

“…Cen, 5S and 45S represent centromere, 5S and 45S rDNAs, respectively (Ocalewicz et al 2003) under the light microscope due to the restrictions of banding techniques. Although G-banding could be shown in some species, this required treatment with specific chemicals, which often resulted in destruction of the chromosome structure (Muravenko et al 2003). The technique developed in this study avoids chromosome destruction induced by G-banding treatment and results in clear bands without any treatment except routine chromosome preparation.…”

“…Chromosome G-banding patterns represent natural conformational features of chromatin (Saitoh and Laemmli 1994;Hoshi and Ushiki 2001;Oberringer et al 2004). In plants, although some reports on G-banding have been published (Drewary 1982;Schubert et al 1984;Murata and Orton 1984;Wang and Kao 1988;Yang and Zhang 1988;Kakeda et al 1990;Peffley and de Vries 1993;Song et al 1994;Muravenko et al 1999Muravenko et al , 2003, a satisfactory Gbanding technique, suitable for different plant species, has not yet been established. In particular, closely adjacent multiple fluorescence bands, including Q-bands and similar have never been resolved.…”