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

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Presentation on theme: "GT training- A GOSH experience Sumera Ghani Great Ormond St Hospital London Paediatric Malignancy Unit."— Presentation transcript:

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

2 About Me Sept July BSc in Human Genetics University College London (UCL) Sep Oct MSc in Experimental Oral Pathology at Queen Mary University of London August 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 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. PMCF- Genetics core facility

10 DNA and RNA extraction from: Blood Bone marrow Fixed material Solid tumour Paraffin embedded material DNA Integrity/concentration

11 RNA AND DNA QUALITY LADDERLADDER DNA1DNA1 DNA2DNA2 DNA3DNA3 RNA1RNA1 RNA2RNA2 RNA3RNA3

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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