Molecular Diagnostics of Cancer 2 Flashcards
(26 cards)
What is RT-PCR?
RT-PCR (Reverse Transcriptase PCR) uses mRNA as a template and converts it to cDNA.
What is qPCR?
qPCR (Quantitative PCR) provides quantification of template using fluorescent probes.
Why do reaction products need to differ in shape and size?
The reaction products need to be different in shape and size in order for them to be told apart.
How can mutations affect PCR products?
Mutations can result in the creation or deletion of a restriction site in a gene, leading to polymorphisms in the size of the PCR product.
What are point mutations?
Point mutations (PM) are single nucleotide changes affecting one biochemical function and can be detected using PCR.
What are Single Nucleotide Polymorphisms (SNPs)?
SNPs are common in populations with a frequency greater than 1%.
How does PCR detect mutations?
PCR differentiates mutations by product size or reaction failure, and RFLPs can arise from mutations that add or remove restriction sites.
What is NSCLC?
Non-Small Cell Lung Cancer (NSCLC) accounts for approximately 80% of all lung cancers.
Where are EGFR gene mutations found?
EGFR gene mutations are found in exons 18, 19, and 21.
How do EGFR mutations affect treatment?
EGFR mutations increase sensitivity to 1st, 2nd, and 3rd generation EGFR TK inhibitors (e.g., gefitinib, erlotinib).
How are EGFR mutations detected?
EGFR mutations are detected through PCR-based methods, such as ASPCR.
What is Allele-Specific PCR (ASPCR)?
ASPCR detects point mutations without restriction sites and relies on precise 3’ end primer matching.
What are the general features of microarray technologies?
Microarray technologies assess gene expression, genotype, or sequence using high-density DNA/RNA probe arrays.
What are the applications of microarray technologies?
Applications include basic research, human diagnostics, and personalized medicine.
What is gene expression profiling?
Gene expression profiling differentiates patterns like up/downregulation in cancer samples.
What is the Affymetrix GeneChip®?
Affymetrix GeneChip® features high-density SNP and copy number variation probes.
What is Next-Generation Sequencing (NGS)?
NGS is massively parallel, high-throughput sequencing that allows rapid, cost-effective gene or genome sequencing.
What is the Illumina sequencing workflow?
The Illumina sequencing workflow includes genomic DNA preparation, fragmentation, adaptor ligation, and amplification.
What occurs during sequencing by synthesis?
Imaging captures the fluorescent signal from incorporated nucleotides, and cyclic chemistry allows for continuous sequencing.
What are the key features of PacBio SMRT sequencing?
PacBio SMRT sequencing includes single molecule sequencing, zero-mode waveguides (ZMWs), and real-time continuous observation.
What are BRCA mutations associated with?
BRCA1/2 mutations are linked to breast and ovarian cancer risk and are involved in the homologous recombination DNA repair pathway.
What is Sanger sequencing?
Sanger sequencing uses dideoxy NTPs as chain terminators, resulting in end-labelled sequences of varying lengths.
What are real-time sequencing technologies?
Real-time sequencing technologies include Oxford Nanopore and Pacific Biosciences.
How does Oxford Nanopore sequencing work?
Oxford Nanopore sequencing threads DNA through a nanopore, detecting changes in electric current as nucleotides pass through.
