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GT training- A GOSH experience

GT training- A GOSH experience. Sumera Ghani Great Ormond St Hospital London Paediatric Malignancy Unit. About Me. Sept. 2003- July 2006- BSc in Human Genetics University College London (UCL) Sep. 2007- Oct. 2008- MSc in Experimental Oral Pathology at Queen Mary University of London

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GT training- A GOSH experience

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  1. GT training- A GOSH experience Sumera Ghani Great Ormond St Hospital London Paediatric Malignancy Unit

  2. About Me • Sept. 2003- July 2006- BSc in Human Genetics University College London (UCL) • Sep. 2007- Oct. 2008- MSc in Experimental Oral Pathology at Queen Mary University of London • August 2008- Started working as Genetic Technologist at Great Ormond St Hospital NHS trust

  3. About the lab • CPA accredited lab • UK NEQAS The team comprises of a: • Consultant Cytogeneticist • Section Head/Principal Molecular Geneticist • Principal Clinical Cytogeneticist • Chief Biomedical Scientist • Senior Clinical Cytogeneticist • 3 Clinical Cytogeneticist • 2 Medical Technical Officers

  4. PMU OVERVIEW . PMCU- Diagnostic Cytogenetics Unit We work closely with the haematology/oncology department and their consultants within GOSH. PMCF- Genetics Core Facility • New facility established in September 2008 • Provide both Molecular Genetics and Cytogenetics tests to: NHS Trusts Academic Institutions Private Labs Pharmaceutical Industries

  5. PMCU- Cytogenetic training • Booking in samples- • Solid tumour • Blood • Bone Marrow • Lymph nodes • Setting up cultures • Harvesting • Slide making • Banding

  6. Techniques Learnt: • G banding • Culturing Solid tumours and preparing slides. • Setting up FISH and analysis • Auditing • Extraction of Mononuclear cells for freezing in liquid Nitrogen • DNA and RNA extraction • Agarose Gel analysis • Setting up a PCR reaction • Sequencing • Growing Fosmids and BACS • Extracting DNA from Fosmids and BACS • Nick translation- labelling DNA

  7. FISH (Fluorescent In Situ Hybridisation) • SETTING UP FISH • Paraffin Embedded Slides • Touch Imprints • Bone Marrow • Solid tumour • Blood • DIFFERENT TYPES • Whole Chromosome Paints • LSI FISH • Metaphase FISH

  8. FISH (Fluorescent In Situ Hybridisation)Locus specific probe t(9;22)(q34;q11).

  9. PMCF- Genetics core facility • In addition to chromosome analysis (karyotyping) • a full range of adjunctive tests, such as Fluorescence in Situ Hybridisation (FISH), • Reverse Transcriptase polymerase chain reaction (RT-PCR) for translocation detection of specific fusion genes. • Mutation detection for a number of genes using direct sequencing, genotyping and MLPA.

  10. PMCF- Genetics core facility • DNA and RNA extraction from: • Blood • Bone marrow • Fixed material • Solid tumour • Paraffin embedded material • DNA Integrity/concentration

  11. RNA AND DNA QUALITY L A D D E R D N A1 D N A2 D N A3 R N A1 R N A 2 R N A3

  12. PCR

  13. Overall experience • Core skills gained in both Molecular and Cytogenetics • Transferable skills • Opportunity to rotate and continue to develop • Opportunity to continue personal learning • Broad experience • Lab project

  14. Questions?

  15. Methods of extracting DNA/RNA • Flexigene- DNA • Trizol/chloroform-RNA • Phenol/chloroform-DNA • Ambion-DNA

  16. PCR

  17. Samples that are accepted for cytogenetic investigations include: • ALL  • AML • JMML/CML  • MDS  • Other leukaemia • TAM • Congenital haematological conditions • Infiltrating tumours • Post-treatment remission samples will be accepted if a prior cytogenetic abnormality has been detected. • Additionally, pleural fluid or CSF diagnostic and relapse specimens may be accepted.

  18. ELA2 NEUTROPHILS

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