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Meaning
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Uses
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MethodSAMPLE PREPARATIONCultured or uncultured lymphocytes are fixed onto a microscope slide.Slide is soaked in SSCSlide is dehydrated in alcohol
Probe is added to the slideProbe area is sealed with a coverslip
DENATURATIONHeat slide to 800C for 5 minutesThe double strands of sample and probe DNA break apart (denature)
HYBRIDISATIONIncubate slide overnight at 370CSpecific probe region will bind to corresponding DNA region
POST HYBRIDISATIONSoak slides in SSC at 640C for 2 minutes - removes non specific, looselybound DNA
Rinse slides in alcoholAdd Dapi/Antifade solutionSeal with a coverslipMICROSCOPYAnalyse metaphases for presence of control regionAnalyse relevant chromosome for probe presence/absenceCount nuclei, noting deletion status
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Temperature controlled slide
processing system
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Slide
Technologist The relevant
excitation filter is
used depending on
the specificwavelength of the
fluorophore being
used. This will
excite the dye on
the slide whichwill emit light that
is filtered by the
emission filter to
give a
homogeneouscolour for each
probe being
examined
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Aneuploidy FISHAnalysis for Aneuploidy
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Aneuploidy
Turner syndrome
Aneuploidy Fish
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Aneuploidy
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Aneuploidy
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Aneuploidy
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No Deletion Found (2 green control signals; 2 red DNA syndrome probe signals)
Deletion Detected (2 green control signals; 1 red DNA syndrome probe signals)
Microdeletion Probes
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Di George
Clinical Symptomsinclude one or a combination of the following:congenital heart disease (particularly conotruncal malformations)palatal abnormalities [especially velopharyngeal insufficiency (VPI)]hypocalcemiaimmune deficiencylearning difficultiescharacteristic facial featuresDiagnosis
FISH detection of a 22q11.2 deletion in the DiGeorge chromosomal region(DGCR).Cytogenetic studies for the presence of a translocation involving 22q11.2(1%)
Deletion 10p13-p14.Mode of InheritanceThe 22q11.2 deletion is inherited in an autosomal dominant manner.About 93% of probands have a de novo deletion of 22q11.2.7% inherited the 22q11.2 deletion from a parent.Both parents of an individual with 22q11.2 deletion should have FISH testingGermline mosaicism
del 22q11.2.Includes:
DiGeorge Syndrome (DGS)
VelocardiofacialSyndrome (VCFS)
Shprintzen Syndrome
Conotruncal Anomaly
Face Syndrome (CTAF)
Caylor Cardiofacial
Syndrome
Autosomal DominantOpitz G/BBB Syndrome
22q11.2 Deletion Syndrome
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22q11.2 Deletion Syndrome
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Microdeletion 15q12
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Microdeletion 15q12Deletion on Paternal Chromosome 15 leads to PW syndrome.
Deletion on Maternal chromosome 15 leads to Angelman Syndrome.
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Microdeletion 15q12
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Whole chromosome PaintUSES:TRANSLOCATIONS
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USES:TRANSLOCATIONS
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USES:TRANSLOCATIONS
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USES:TRANSLOCATIONS
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Preimplantation Genetics
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Preimplantation Genetics
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Preimplantation Genetics
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USES: ONCOLOGY
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USES: ONCOLOGY
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USES: ONCOLOGY
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Software measures HER2 amplification in breast tumor. For eachcell the software reports HER2 & CEP17 copy numbers and the
HER2:CEP17 ratio.