Services and Instrumentation
Cytogenetics
- Study of genetic instability by chromosome analysis
- Cell line identification: human, mouse, rat, dog, monkey or of any other mammalian origin
- Giemsa (G), C, Q, R –banding, SCE (Sister Chromatid Exchange) and silver staining of metaphase chromosomes
- Analysis of meiotic chromosomes of laboratory animals
- Effects of pharmaceutical products on human and other mammalian chromosomes
- Fluorescence in situ hybridization (FISH) on interphase and metaphase cells, paraffin sections, tissue microarray and frozen sections using whole chromosome paints, centromere probes, gene mapping with commercial and laboratory-generated DNA probes
- Quantitative FISH (Q-FISH) analysis of telomeric sequences
- Spectral karyotyping (SKY) of human, mouse and rat cells
- Determination of the genetic suitability of mammalian stem cells to be used for cloning purposes
STR Fingerprinting Service
Short Tandem Repeat (STR) DNA profiling, also known as DNA fingerprinting, offers the greatest value for cell line authentication. The assay is based on screening regions of microsatellite instability with defined tri- or tetrad-nucleotide repeats located throughout the chromosomes. PCR reactions using primers on non-repetitive flanking those regions will generate PCR products of different sizes based on the number of repeats in the region; the size of these PCR products is determined by capillary electrophoresis. By combining between 8 and 16 STR loci, such as D5S818, D13S317, D7S820, D16S539, vWA, TH01, TPOX, and CSF1PO, it is possible to uniquely identify a sample. The CCLC assay screens 16 loci using the Promega Powerplex 16 HS kit. Our test also includes matching the STR profiles against an internal database comprised of public profiles and profiles that are unique to cell lines developed by MDACC investigators. Our current database has over 4000 profiles.
Mycoplasma Contamination Testing
Mycoplasma is a bacterial contaminant commonly found in cell culture. It is easy to spread from culture to culture, and unlike most other bacterial or fungal contaminants, mycoplasma contamination cannot be easily detected without further testing. Mycoplasma are bacteria that do not contain a traditional cell membrane and can alter the metabolic characteristics of cells in culture negatively affecting experimental results. We provide this mycoplasma determination service using the Lonza MycoAlert Kit.
Purchase Cell Lines
The CCLC has a List of Cell Lines that have been validated and tested for mycoplasma contamination that are available for purchase.
Instrumentation
CCAC is equipped with:
- Nikon 80i microscope with 10X, 20X, and 63X plan Apo objectives
- Nikon 80i microscope with 10X, 20X, and 60X plan Apo objectives
- Nikon Eclipse 400 microscope with 10X and 63X objectives
- Nikon Eclipse 200 microscope with 10X objective
- Nikon 50i microscope with 10X, 60X and 100X objectives
- Nikon Eclipse TS100 microscope with 10X and 40X objectives
- Sensys 1060x540 CCD camera
- Two Applied Spectral Imaging karyotyping workstations (SKY) and spectral karyotyping interferometer plus workstations.
- State of the Art Applied Spectral Imaging Hyperspectral Karyotyping software.
- CoolSNAP HQ2 Monochrome camera: 14 bit, 20MHz Digital monochrome Camera, Motorized focus control with Z stacking capability
- NIS-Elements: Advanced Research Acquisition and analysis package