Chromosome Banding Techniques Research Articles

Chromosomal Characterization of Five Medicinally Significant Phyllanthus Species in Bangladesh by DNA Base-Specific Fluorochrome Banding Technique

ABSTRACT Five Phyllanthus species were characterized using a fluorochrome chromosome banding technique with CMA and DAPI. This genus displayed a range of somatic chromosomal counts, including 2n = 2x = 26 (diploid) in P. acidus, P. niruri, and P. reticulatus; 2n = 6x = 48 (hexaploid) in P. urinaria; and 2n = 10x = 100 (decaploid) in P. emblica (wild and cultivated varieties), which exhibited a multi-basic chromosome number. The centromeric and terminal regions of the chromosomes contained most of the CMA and DAPI bands, indicating that GC- and AT-rich repeats had accumulated in these locations. The diversity based on the heterochromatin distribution patterns made it possible to use the CMA and DAPI banding approaches to analyze and characterize these five Phyllanthus species.

A Protocol for the Detection of Fusion Transcripts Using RNA-Sequencing Data.

Fusion transcripts are formed when two genes or their mRNAs fuse to produce a novel gene or chimeric transcript. Fusion genes are well-known cancer biomarkers used for cancer diagnosis and as therapeutic targets. Gene fusions are also found in normal physiology and lead to the evolution of novel genes that contribute to better survival and adaptation for an organism. Various in vitro approaches, such as FISH, PCR, RT-PCR, and chromosome banding techniques, have been used to detect gene fusion. However, all these approaches have low resolution and throughput. Due to the development of high-throughput next-generation sequencing technologies, the detection of fusion transcript becomes feasible using whole genome sequencing, RNA-Seq data, and bioinformatics tools. This chapter will overview the general computational protocol for fusion transcript detection from RNA-sequencing datasets.

Citogenética comparativa em felídeos (Carnivora: Felidae): Leopardus wiedii, Panthera onca, Puma concolor e Felis catus

ABSTRACT This study aims to conduct a cytogenetic analysis on four South American feline species: the jaguar (Panthera onca), the puma (Puma concolor), the ocelot (Leopardus wiedii), and the domestic cat (Felis catus). We discovered that the jaguar and the puma mainly differ in their fundamental number of chromosomes. Furthermore, we observed a morphological difference in a pair of chromosomes between the jaguar and the domestic cat. This suggests that, over the course of evolution, there was a pericentric inversion in a pair of B4 group chromosomes, leading them to become part of the A group in the jaguar. When analyzing the puma, we noticed the presence of a fourth pair of chromosomes in the E group, along with a deletion in the F group. Additionally, we observed the presence of an additional pair of chromosomes in the C group in the ocelot. Another interesting aspect of the study was the location of nucleolar organizer regions (NORs) on the chromosomes. To elucidate the origin of other chromosome pairs, we employed chromosomal banding techniques, allowing us to better understand the evolution of the C3 pair in the Leopardus genus and the E4 pair in the Puma genus.

Molecular Cytogenetics in Domestic Bovids: A Review.

The discovery of the Robertsonian translocation (rob) involving cattle chromosomes 1 and 29 and the demonstration of its deleterious effects on fertility focused the interest of many scientific groups on using chromosome banding techniques to reveal chromosome abnormalities and verify their effects on fertility in domestic animals. At the same time, comparative banding studies among various species of domestic or wild animals were found useful for delineating chromosome evolution among species. The advent of molecular cytogenetics, particularly the use of fluorescence in situ hybridization (FISH), has allowed a deeper investigation of the chromosomes of domestic animals through: (a) the physical mapping of specific DNA sequences on chromosome regions; (b) the use of specific chromosome markers for the identification of the chromosomes or chromosome regions involved in chromosome abnormalities, especially when poor banding patterns are produced; (c) better anchoring of radiation hybrid and genetic maps to specific chromosome regions; (d) better comparisons of related and unrelated species by comparative FISH mapping and/or Zoo-FISH techniques; (e) the study of meiotic segregation, especially by sperm-FISH, in some chromosome abnormalities; (f) better demonstration of conserved or lost DNA sequences in chromosome abnormalities; (g) the use of informatic and genomic reconstructions, in addition to CGH arrays, to predict conserved or lost chromosome regions in related species; and (h) the study of some chromosome abnormalities and genomic stability using PCR applications. This review summarizes the most important applications of molecular cytogenetics in domestic bovids, with an emphasis on FISH mapping applications.

100 Years of Chromosome Research in Rye, Secale L.

Although microscopy and genetics were still in their infancy, there are cytological results produced a hundred years ago that are still relevant today. Since the 1920s, rye has been a subject of chromosome research. It started by plotting its mitotic and meiotic chromosomes to determine genome size. After controversial evidence, it became clear that the base number is n = 7. However, structural differences exist between species within the genus Secale. Some rye populations even carry accessory chromosomes evolutionary derived from the A genome. The development of tetraploid strains significantly promoted chromosome analysis. Various techniques have tried to stabilize the disturbed chromosome pairing of the induced tetraploids. Although slight improvements could be achieved, they did not lead to a breakthrough. However, the various aneuploid derivatives of the polyploids found major advances in the genetic analysis of rye. Trisomics, telo-trisomics, and reciprocal translocation have served as important tools for gene mapping. Since the 1970s, various chromosome banding techniques have stimulated scientific progress. The seven haploid chromosomes could be diagnosed unequivocally, not only in S. cereale but also in related species. These findings led to a clear homoeologous assignment to the genomes of related grass species such as wheat, barley, rice, etc. Current applications of in situ fluorescence staining methods, such as GISH and FISH, allow even more precise results, depending on the specificity of the DNA samples. Advanced preparation techniques are supplemented by the variety of innovations in the field of molecular genome analysis. They replace complex cytological examinations. In this way, introgressions can be safely detected by DNA markers and be much more detailed. In addition, CRISPR/CAS-mediated chromosome engineering will become an important method of the future.

Placing women in Cytogenetics: Lore Zech and the chromosome banding technique

BackgroundScholars agree that Torbjörn Caspersson’s lab at the Institute of Medical Cell Research and Genetics at the Karolinska Institute, Sweden, played a key role in the first description of the so-called Q-banding technique. It laid the foundation for a new era of cytogenetic diagnostics and had a lasting impact in several areas of biology and medicine.MethodsBased on a mixed-method approach, essential aspects of the history of human cytogenetics are considered via primary and secondary analysis of biographical interviews as well as the qualitative evaluation of bibliometrics. Drawing on interviews with colleagues of lab member Lore Zech (1923–2013) and contemporary publications, this paper illuminates the role of and contribution by Zech: To what extent is the discovery attached to her and what does her legacy look like today?ResultsThe analysis of the contemporary witness interviews with colleagues, students and junior researchers shows that Lore Zech was a committed member of Caspersson's research group. In addition, memoirs by contemporary colleagues describe her outstanding skills in microscopy. The different sources paint a multifaceted picture. In addition to the historians' patterns of interpretation, different legacies can also be found within the peer group.ConclusionsWe argue that Zech represent the type of scientist who, although her research was acknowledged with several prizes, so far has not been part of the canon of pioneers of international cytogenetics.

Clinical Cytogenetics of the Dog: A Review.

Simple SummaryThe cytogenetic analysis of dogs is mainly focused on the diagnosis of disorders of sex development (DSD) and cancers. Unfortunately, the study of canine chromosomes is a challenging task due a high chromosome number (2n = 78) and the one-arm morphology of all autosomes. For years, the application of conventional cytogenetic techniques, Giemsa staining and G and DAPI (4′,6-diamidino-2-phenylindole) bandings, allowed the identification of sex chromosome aneuploidies and centric fusions. An advanced clinical cytogenetic diagnosis is also needed due to the fact that the dog is a valuable animal model in biomedical research. The application of hybridization methods, such as fluorescence in situ hybridization (FISH) and array comparative genome hybridization (aCGH), facilitated the detection of other chromosomal rearrangements. It can be foreseen that a wide use of modern molecular techniques (e.g., SNP microarray and next generation sequencing) will substantially extend the knowledge on canine chromosome mutations.The dog is an important companion animal and has been recognized as a model in biomedical research. Its karyotype is characterized by a high chromosome number (2n = 78) and by the presence of one-arm autosomes, which are mostly small in size. This makes the dog a difficult subject for cytogenetic studies. However, there are some chromosome abnormalities that can be easily identified, such as sex chromosome aneuploidies, XX/XY leukocyte chimerism, and centric fusions (Robertsonian translocations). Fluorescence in situ hybridization (FISH) with the use of whole-chromosome painting or locus-specific probes has improved our ability to identify and characterize chromosomal abnormalities, including reciprocal translocations. The evaluation of sex chromosome complement is an important diagnostic step in dogs with disorders of sex development (DSD). In such cases, FISH can detect the copy number variants (CNVs) associated with the DSD phenotype. Since cancers are frequently diagnosed in dogs, cytogenetic evaluation of tumors has also been undertaken and specific chromosome mutations for some cancers have been reported. However, the study of meiotic, gamete, and embryo chromosomes is not very advanced. Knowledge of canine genome organization and new molecular tools, such as aCGH (array comparative genome hybridization), SNP (single nucleotide polymorphism) microarray, and ddPCR (droplet digital PCR) allow the identification of chromosomal rearrangements. It is anticipated that the comprehensive use of chromosome banding, FISH, and molecular techniques will substantially improve the diagnosis of chromosome abnormalities in dogs.

Chromosome Abnormalities and Fertility in Domestic Bovids: A Review.

Simple SummaryIn domestic bovids, numerical autosome abnormalities have been rarely reported, as they present abnormal animal phenotypes quickly eliminated by breeders. However, numerical abnormalities involving sex chromosomes and structural (balanced) chromosome anomalies have been more frequently detected because they are most often not phenotypically visible to breeders. For this reason, these chromosome abnormalities, without a cytogenetic control, escape animal selection, with subsequent deleterious effects on fertility, especially in female carriers.After discovering the Robertsonian translocation rob(1;29) in Swedish red cattle and demonstrating its harmful effect on fertility, the cytogenetics applied to domestic animals have been widely expanded in many laboratories in order to find relationships between chromosome abnormalities and their phenotypic effects on animal production. Numerical abnormalities involving autosomes have been rarely reported, as they present abnormal animal phenotypes quickly eliminated by breeders. In contrast, numerical sex chromosome abnormalities and structural chromosome anomalies have been more frequently detected in domestic bovids because they are often not phenotypically visible to breeders. For this reason, these chromosome abnormalities, without a cytogenetic control, escape selection, with subsequent harmful effects on fertility, especially in female carriers. Chromosome abnormalities can also be easily spread through the offspring, especially when using artificial insemination. The advent of chromosome banding and FISH-mapping techniques with specific molecular markers (or chromosome-painting probes) has led to the development of powerful tools for cytogeneticists in their daily work. With these tools, they can identify the chromosomes involved in abnormalities, even when the banding pattern resolution is low (as has been the case in many published papers, especially in the past). Indeed, clinical cytogenetics remains an essential step in the genetic improvement of livestock.

Cytogenetic Investigations in Emballonuroidea. II. Chromosome Painting in Nycteridae Reveals Cytogenetic Signatures Pointing to Common Ancestry of Nycteris and Emballonura

Two species of the monogeneric family Nycteridae were studied for the first time with chromosome banding techniques and chromosome painting. The diploid chromosome number of Nycteris macrotis and N. tragata is 40 and 38, respectively. Both karyotypes differ by a translocation, a telomeric fusion and a pericentric inversion. The genus Nycteris shows a highly derived karyotype where the 25 chiropteran evolutionarily conserved units (ECUs) are represented in 37 segments. Chromosome painting with Myotis probes revealed three common features of Nycteris and Emballonura, which are not present in Taphozous. From a cytogenetic point of view, Nycteris is therefore closer related to Emballonura than to Taphozous. Further, an additional synapomorphy for Vespertilioniformes is described, i.e. the synteny of a segment homologous to human chromosome 13 and ECU12a-22a in elements homologous to Myotis chromosome MMY6.

Karyotype characterization and evolution of chromosome number in Cactaceae with special emphasis on subfamily Cactoideae

ABSTRACT Cactaceae species are karyotypically well-known with x = 11 and chromosome number variation due mainly to polyploidization. However, both assumptions are based on descriptive observations without taking an evolutionary framework of Cactaceae into account. Aiming to confirm these hypotheses in an evolutionary context, we obtained chromosome numbers for 20 species of Cactoideae, performed an extensive review of chromosome number for the family, and analyzed these data using a phylogenetic approach. The karyotypes presented here were characterized by CMA/DAPI banding, and for six species 5S and 45S rDNA sites were located. Our data, along with a survey of the literature, reinforce the long-standing hypothesis of a x = 11 as the base chromosome number for Cactaceae. They also reinforce the relevance of polyploidy in karyotype evolution of cacti, although polyploidy was important just after the diversification of subfamilies Maihuenioideae and Pereskioideae. Despite the homogeneous chromosome complements observed among cacti, chromosome banding and FISH techniques revealed informative characteristics, allowing the identification of chromosome synapomorphies, such as proximal CMA+ bands in Melocactus and proximal 5S rDNA in Pilosocereus, indicating the taxonomic potential of chromosome characterization in cacti.

Banded karyotype of Nelore cattle (Bos taurus indicus Linnaeus, 1758).

Chromosome banding techniques were applied and standardized to obtain karyotype characteristics for the first time in Brazil of Nelore cattle – Bos taurus indicus Linnaeus, 1758 – (bovine subspecies most prominent in Brazilian livestock). Blood samples were collected from the animals of the School of Agrarian and Biological Sciences of the Pontifical Catholic University of Goiás, two males and two females of pure breed. These samples were submitted to the cell culture method to study metaphase chromosomes. Chromosome banding techniques (C, G and NOR) revealed the karyotype architecture of Nelore cattle common with that of other breeds of zebu cattle formerly karyotyped. The diploid chromosome number was invariably normal, 2n = 60. C-banding revealed C-positive heterochromatin in centromeric regions almost in all chromosomes. G-banding presented the expected band pattern in the respective chromosome pairs in correspondence with the established chromosomal patterns for the species. Ag-staining for nucleolus organizer regions (AgNOR) was identified on the telomeric end of the long arm in 7 autosomal chromosomes. In this study we found more regions in chromosomes with staining than presented in the literature for the Bos indicus group (BIN). These NOR regions were repeated on the same chromosomes for the 4 animals studied.

Cytological markers used for identification and transfer of Aegilops spp. chromatin carrying valuable genes into cultivated forms of Triticum.

There are many reports describing chromosome structure, organization and evolution within goatgrasses (Aegilops spp.). Chromosome banding and fluorescence in situ hybridization techniques are main methods used to identify Aegilops Linnaeus, 1753 chromosomes. These data have essential value considering the close genetic and genomic relationship of goatgrasses with wheat (Triticumaestivum Linnaeus, 1753) and triticale (× Triticosecale Wittmack, 1899). A key question is whether those protocols are useful and effective for tracking Aegilops chromosomes or chromosome segments in genetic background of cultivated cereals. This article is a review of scientific reports describing chromosome identification methods, which were applied for development of prebreeding plant material and for transfer of desirable traits into Triticum Linnaeus, 1753 cultivated species. Moreover, this paper is a resume of the most efficient cytomolecular markers, which can be used to follow the introgression of Aegilops chromatin during the breeding process.

Reciprocal Translocation Analysis with Whole Chromosome Painting for FISH.

Ionizing radiation can produce many chromosome aberrations; however, a number of aberrations are not readily detectable with a conventional Giemsa staining and require special staining methods. Moreover, an easy method to identify nondetectable chromosomal aberrations, such as symmetrical inter-chromosomal translocations, is available: whole chromosome fluorescence in situ hybridization (FISH) staining. Asymmetrical translocations such as dicentric chromosomes are known to be lethal but cells with symmetrical translocations may survive and persist in the progeny population for years. Chromosome banding techniques can also detect inter-chromosomal translocations but understanding banding patterns requires professional training and experience. Due to its clear detection by a discernible painting color, whole chromosome painting method is superior in identifying chromosome fragmentation, loss, and amplification of chromosomes. This chapter introduces whole chromosome FISH painting method to detect chromosomal aberrations such asfragments and translocations.

Cytogenetic Trends in Two Families of the Neotropical Catfishes: Heptapteridae and Pseudopimelodidae (Siluriformes).

Several neotropical Siluriformes groups suffered important taxonomic revisions based on the evaluation of morphological and molecular characteristics that allow the construction of new phylogenetic hypothesis. In the present study were cytogenetically analyzed six species belonging to Heptapteridae (Cetopsorhamdia iheringi, Phenacorhamdia tenebrosa, Rhamdella eriarcha, Pimelodella meeki, Pimelodella australis, Heptapterus mustelinus) and two to Pseudopimelodidae families (Microglanis cottoides and Microglanis cibelae) by means of differential staining techniques to describe more precisely cytogenetic similarities and differences. The diploid number of R. eriarcha with 2n = 58 and M. cibelae with 2n = 56 were reported for the first time. Also, the lowest chromosome number (2n = 48) for P. tenebrosa was described. The chromosome-banding techniques for to put in evidence nucleolar organizers impregnated by silver nitrate ([AgNORs], chromomycin A3 [CMA3], and rDNA 18S) showed for all studied species conserved patterns, characteristic for each family. The rDNA 5S showed high variability among species or populations of both families, these regions could be simple or multiple, syntenic, or not with rDNA18S. The chromosome markers showed that both families are related not only from a morphologic point of view but also by their karyotypic characteristics, however, some of the present cytogenetic results evidence the importance of new morphologic, molecular, and phylogenetic studies to improve the knowledge of these fish groups.

Use of Array Comparative Genomic Hybridization for the Diagnosis of DiGeorge Syndrome in Saudi Arabian Population

DiGeorge syndrome (DGS) is a genetic disorder known as a clinically variable syndrome with over 180 associated phenotypic features. It is caused by a common human deletion in the 22q11.2 chromosomal region and currently is affecting approximately 1 in 4,000 individuals. Despite the prevalence of inherited diseases mainly due to consanguineous marriages, the current diagnosis of DGS in Saudi Arabia is mainly based on conventional high-resolution chromosome banding (karyotyping) and FISH techniques. However, advanced genome-wide studies for detecting microdeletions or duplications across the whole genome are needed. The aim of this study is to implement and use aCGH technology in clinical diagnosis of the 22q11.2 deletion in Saudi Arabian DGS patients and to confirm its effectiveness compared to conventional FISH and chromosome banding techniques. Thirty suspected DGS patients were assessed for chromosome 22q11.2 deletion using high-resolution G-banding, FISH, and aCGH. The aCGH results were compared with those obtained by the other 2 cytogenetic techniques. G-banding detected the 22q11.2 deletion in only 1 patient in the cohort. Moreover, it detected additional chromosomal aberrations in 3 other patients. Using FISH, allowed for detection of the 22q11.2 deletion in 2 out of 30 patients. Interestingly, the use of aCGH technique showed deletions in the chromosome 22q11.2 region in 8 patients, indicating a 4-fold increase in diagnostic detection capacity compared to FISH. Our results show the effectiveness of aCGH to overcome the limitations of FISH and G-banding in terms of diagnostic yield and allow whole genome screening and detection of a larger number of deletions and/or duplications in Saudi Arabian DGS patients. Except for balanced translocations and inversions, our data demonstrate the suitability of aCGH in the diagnostics of submicroscopic deletion syndromes such as DGS and most chromosomal aberrations or complex abnormalities scattered throughout the human genome. Our results recommend the implementation of aCGH in clinical genomic testing in Saudi Arabia to improve the diagnostic capabilities of health services while maintaining the use of conventional cytogenetic techniques for subsequent validation or for specific and known aberrations whenever required.

Karyology of Agapanthus africanus (L.) Hoffmanns (Agapanthaceae)

The genus Agapanthus L'Héritier (Agapanthaceae) is a monocotyledonous, herbaceus and perennial plant group endemic to southern Africa. Ten species have been described for this group, in which three are considered evergreen and seven deciduous plants. Nevertheless, Agapanthus L'Héritier taxonomy is considered very difficult once there are few unique characters and a high plasticity depending on circumstances of growing. This feature takes some authors to believe in the existence of only one species with innumerous varieties.The cytogenetic has been an important tool to aid the plant taxon identification, and to date, all taxa of Agapanthus L'Héritier studied cytologically presented 2n = 30. Although the species possess large chromosomes, few karyological studies has been explored. This work aimed to increase the cytogenetic knowledge of Agapanthus africanus (L.) Hoffmanns by chromosome banding techniques with DAPI / CMA3 and Fluorescent in situ Hybridization (FISH). In addition, flow cytometry was used to determinate DNA content and the percentage of AT / GC nitrogenous bases. Plants studied showed 2n = 30 chromosomes, ranging from 4.34 - 8.55 µm, with karyotype formulae (KF) = 10m + 5sm. The FISH technique revealed one 45S rDNA signal observed proximally to centromere of the chromosome 7, and one 5S rDNA signal observed proximally to centromere of chromosome 9. The 2C DNA content estimated for the species was 2C = 24.4pg, with 59% of AT and 41% of GC. Our data allowed important upgrade for biology and cytotaxonomy of Agapanthus africanus (L.) Hoffmanns, and will be used to future taxonomic and phylogenetics studies, as well as for the chromosome evolution of the genus. APOIO FINANCEIRO: Fapemig, Capes, CNPq

Chromosome Banding in Amphibia. XXXV. Highly Mobile Nucleolus Organizing Regions in Craugastor fitzingeri (Anura, Craugastoridae)

A 7-year cytogenetic study on the leaf litter frog Craugastor fitzingeri from Costa Rica and Panama revealed the existence of highly mobile nucleolus organizing regions (NORs) in their genomes. Silver (Ag)-staining of the active NORs demonstrated an exceptional interindividual pattern of NOR distribution at the telomeres of the chromosomes. All individuals examined showed a different and specific NOR location in their karyotypes. Furthermore, intraindividual variation in the NOR sites was found. This observation suggested the existence of mobile NORs in C. fitzingeri. Confirmation of this phenomenon was possible by systematic FISH analysis using an 18S + 28S rDNA probe. The extremely variable number and position of the NORs in C. fitzingeri is best explained by highly mobile NORs that move freely between the telomeres of the chromosomes. These transpositions must occur preferentially in premeiotic, meiotic, or postmeiotic stages, but also at a lower incidence in the somatic tissues of the animals. It is hypothesized that transposable (mobile) elements are closely linked to the NORs or are inserted into the major 18S + 28S rDNA spacers of C. fitzingeri. When such transposable elements spread by transpositions, they can carry with them complete or partial NORs. The present study provides detailed information on various differential chromosome banding techniques, in situ hybridization experiments, chromosomal hypermethylation patterns, determination of the genome size, and analyses of restriction fragment length polymorphisms of the DNA.

Estudios citogenéticos en cuatro taxones de Canna (Cannaceae) de la provincia de Buenos Aires, Argentina

Se investigaron los números cromosómicos y los cariotipos de tres especies y un híbrido del género Canna, que crecen en la Argentina y también se encuentran distribuidos en el neotrópico. Mediante una tinción cromosómica convencional se analizaron C. indica, C. fuchsina, C. coccinea y el híbrido C. x generalis. Todos los taxones estudiados resultaron diploides (2n = 18). Las tres especies presentaron cariotipos similares simétricos, con cromosomas pequeños, mayormente metacéntricos aunque con algunos pares cromosómicos submetacéntricos. En C. coccinea y C. fuchsina, y por primera vez en el género, se aplicó la técnica de bandeo fluorescente DAPI sobre interfases y metafases mitóticas, con el objetivo de detectar regiones de heterocromatina constitutiva rica en Adenina-Timina (A-T). Los resultados obtenidos permitieron el reconocimiento de pares cromosómicos y demostraron la utilidad de este método para detectar la variación interespecífica en la distribución cromosómica de la heterocromatina rica en A-T en Canna. Se sugiere que el material estudiado de C. x generalis es un diploide en el que el 2n = 18 no contradice la hipótesis sobre su origen, ya que el número hallado coincide con el de uno de los progenitores, C. indica, y con el que se conoce para otras especies parentales.

Chromosome Bandings.

Chromosome banding is an essential technique used in chromosome karyotyping to identify normal and abnormal chromosomes for clinical and research purposes. Giemsa (G)-, reverse (R)-, and centromere (C)-banding are the most commonly dye-based chromosome-banding techniques. G-banding involves the staining of trypsin-treated chromosomes and R-banding involves denaturing in hot acidic saline followed by Giemsa staining. C-banding is specifically used for identifying heterochromatin by denaturing chromosomes in a saturated alkaline solution followed by Giemsa staining. Different banding techniques may be selected for the identification of chromosomes.

Cytogenetic analysis for research and services

Abstract That the correct chromosome number in man is 46 was first recognized by Tjio and Levan in 1956. Perhaps few Indonesians know that Tjio was an Indonesian scientist studying in Sweden and then living in the US. Cytogenetic analyses are commonly performed to determine both structural and numerical chromosome aberration, whilst changes in chromosomes can lead to birth defects, syndromes, or even cancer. Several chromosomal aneuploidy syndromes were identified after the establishment of various chromosome banding techniques in late 1960’s. Specific cell culture media was found to express fragile site in the beginning of 1970’s and since then, inherited Fragile X Mental Retardation syndrome could be diagnosed. However, some female permutation cases have been often misdiagnosed. Further molecular analysis has resolved this problem by revealing more CGG repeats in the promoter region FMR1 gene, which is related to the expression of fragile site and the severity of the diseases. In Disorder of Sex Development (DSD), early gender assignment and reconstruction surgery has been challenged because of the dilemma of gender identity development in later life. Cytogenetic analysis for the first-line gender assignment is important in newborn with DSD. Proper diagnosis with hormonal and mutation analysis should be elucidated to avoid medical, psychological, and social aspect in adult life. The most frequent genetic cases in our clinical experiences have been Androgen Insensitivity Syndrome and Congenital Adrenal Hyperplasia. Female Complete Androgen Insensitivity Syndrome (CAIS) with main symptom primary amenorrhea without cytogenetic analysis has often been diagnosed as inguinal hernia because of testicle location and size. Diagnosis and treatment of several leukemias and lymphomas, as well as some solid tumors, depend on cytogenetic analyses to demonstrate consistent, specific chromosomal aberrations. Chromosome analysis in hematologic malignancy is indicated to support diagnosis, select therapy regimen, and elaborate prognosis. Specific chromosome translocations have been identified for hematologic malignancy. The breakpoints of several of these translocations have been cloned. Several loci of oncogene have been identified and sequenced. Molecular genetic analysis will replace cytogenetic analysis and shift the requirement for studying metaphase cells. Therefore, chromosome analysis in genetic disease and cancer should be attained with advanced molecular techniques, such as Fluorescence In Situ Hybridization (FISH) and microarray CGH analysis. Cytogenetic analysis is still useful and applicable in genetic disease diagnosis, sexual assignment, and hematologic malignancy in the laboratory with minimal equipments. Molecular analysis as a part of health care services in Indonesia has been limited in research centers in university setting; therefore, a comprehensive diagnosis with genetic analysis has often been improbable.