Gt training a gosh experience
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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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Gt training a gosh experience

GT training- A GOSH experience

Sumera Ghani

Great Ormond St Hospital London

Paediatric Malignancy Unit


About me

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


About the lab

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


Pmu overview

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


Pmcu cytogenetic training

PMCU- Cytogenetic training

  • Booking in samples-

  • Solid tumour

  • Blood

  • Bone Marrow

  • Lymph nodes

  • Setting up cultures

  • Harvesting

  • Slide making

  • Banding


Techniques learnt

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


Fish fluorescent in situ hybridisation

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


Fish fluorescent in situ hybridisation locus specific probe

FISH (Fluorescent In Situ Hybridisation)Locus specific probe

t(9;22)(q34;q11).


Gt training a gosh experience

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.


Pmcf genetics core facility

PMCF- Genetics core facility

  • DNA and RNA extraction from:

  • Blood

  • Bone marrow

  • Fixed material

  • Solid tumour

  • Paraffin embedded material

  • DNA Integrity/concentration


Rna and dna quality

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


Gt training a gosh experience

PCR


Overall experience

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


Questions

Questions?


Methods of extracting dna rna

Methods of extracting DNA/RNA

  • Flexigene- DNA

  • Trizol/chloroform-RNA

  • Phenol/chloroform-DNA

  • Ambion-DNA


Gt training a gosh experience

PCR


Samples that are accepted for cytogenetic investigations include

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.


Ela2 neutrophils

ELA2 NEUTROPHILS


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