The Molecular Cytogenetic Core (MC) at the Albert Einstein College of Medicine provides tools for the preparation of human and murine samples suitable for molecular genetic and cytogenetic analysis of the entire genome. These tools include the establishment of EBV transformed cell lines; isolation of DNA and mRNA from a variety of tissue culture samples as well as primary biopsies; preparation of metaphase chromosomes suitable for fluorescence in situ hybridization (FISH) and Spectral Karyotyping (SKY) or whole chromosome paints for human and mouse genome. The core personnel is trained to hybridize commercial probes and to designed locus specific probes for regions of interest to investigators. All the probes are custom designed and in house generated.
Density gradient centrifugation is used for PBMC isolation. The cell recovery viability is routinely 92%. PBMCs are stored in liquid nitrogen and serum will be kept at -80°C.
Whole genome amplification (WGA) uses DNA polymerase and random primers to amplify the entire genome. It can generate a large amount of DNA directly from a small sample. The core is using the REPLI-g whole genome amplification kit (Qiagen) for large scale and GenomiPhi DNA amplification Kit for small scale WGA.
Qiagen RNeasy Mini Kit is used in the core for RNA isolation from whole blood, cell lines and tissues. We guarantee RNA samples of good quality and free of genomic DNA contamination or RNA degradation.
The core is using Puregene Genomic DNA purification kit, Autogene 610L and traditional Phenol-Chloroform extraction methods to isolate DNA from whole blood, buffy coats, saliva, cell lines, buccal cells, tissues and serum. The 260/280 ratio is routinely of high quality of >1.7.
Epstein Barr Virus (EBV)-transformed human B-lymphoblastoid cell lines.
EBV infection in vitro causes transformation of B cells and generates B lymphoblastoid, permanent cell lines (LCLs). LCLs have been widely used for diagnosis of metabolic disorders. LCLs also have been used as models in various biological and medical studies as they can be easily established and continue proliferating for many generations to provide unlimited DNA/RNA. LCLs will be stored in liquid nitrogen. Our success rate is 90%.
Generation of hamster-human somatic cell hybrid cell lines
Somatic cell hybrids are formed by fusion of cells. Usually the fusion is achieved by polyethylene glycol (PEG) or a virus. After fusion, the new cell has temporarily a double chromosome complement that is gradually reduced by random expulsion of either hamster or human chromosomes. The remaining cells can be tested for the presence of a variety of gene products. The presence, or deficiency, of these products can be associated with the presence, or absence, of a particular chromosome, thus localizing function to site.
The MC is equipped to perform human and mouse SKY. SKY is a powerful 24-color, whole-chromosome genome wide painting assay that can detect chromosomal material of unknown origin, complex rearrangements, translocations, large deletions, duplications and aneuploidy.
The MC offers services for both locus specific probes (BAC clones, plasmids > 5Kb) as well as chromosome painting probes (both human and mouse). We offer the possibility of selecting and ordering BAC clones for any genomic region based on the latest build of the genome. Probes can be generated in up to four colors (Spectrum Orange, Spectrum Aqua, Biotin- Cy5, Digoxigenin –Alexa 488 or Spectrum Green.
Molecular Cytogenetic Core has not received any reviews.
Molecular Cytogenetic Core has not received any endorsements.