Neoplasia molecular pathology: Gene mutations in tumour
9 important questions on Neoplasia molecular pathology: Gene mutations in tumour
What is molecular pathology?
- Detection of genomic alterations in tumors: mutations, translocations, copy number alterations
- Diagnostics
- Differential diagnostics: maligne tumor / reactive proces
- classification of tumors: supporting evidence for tumor type
on cases in which the histological diagnose is inconclusive
- prediction of therapy response based on the tumorgenotype
How are cancer cells killed in targeted therapy?
Difference chemotherapy and targeted therapy
Targeted therapy: focuses on killing cancer cells while doing less damage to normal cells
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Lung cancer and EGFR mutations
Genomic aberrations (afwijkingen) in cancer that are EGFR related are?
- Growthfactor receptor
- Signal transduction
- Transcription factors
- Cell cycle
- Apoptosis
What is Sanger sequencing?
also called "DiDeoxysequencing.“
In this method, nucleotides (A, C, T, G) and terminating nucleotides are incorporated into the synthesized DNA chain during sequence response. As a result, DNA chains of different
lengths are synthesized in sequence response of a DNA template. The DNA chains of different lengths are separated by electrophoresis and
analyzed after completion of the sequence reaction. The signal detected results in a pattern
of peaks
What is next generation sequencing NGS?
NGS also called massive parallel sequencing, in which DNA fragments of different genes and gene regions are generated at the same time.
The technology is based on the "sequencing by synthesis" principle. In the synthesized strand, nucleotides (A, C, T, G) are incorporated during the
sequence reaction. Each nucleotide that is incorporated gives a unique signal detected during the synthesis reaction, hence the term "sequencing by synthesis". Because the individually synthesized DNA chains, reads, are detected, this technique is very sensitive.
What is the working order of Next generations sequencing
- Library formation - PCR
- Amplification
- sequencing
- analysis
Annealing of the adapter-flanked PCR amplicons
- To the ion sphere (use of the complementary P1 sequence)
- clonal amplification
- isolate positive Ion Spheres
- sequencing
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