Chapter 4: Tools of Human Molecular Genetics Flashcards
Technique that:
- uses DNA
- soak gel in NaOH to denature
- radioactive DNA probe
Southern Blot
Technique which:
- matches the normal or the mutation sequence
- detects mutations with 1 base affected or a small # of bases
- only for tested or known mutations
Allele Specific Oligonucleotide
Inherited pair has 1 maternal and 1 paternal chromosome pair
Allelic Exclusion
Technique which:
- is for RNA, using size and abundance
- it gets the cell, separates it by size, denatures, gels it, hybridize, wash and visualize
Northern Blot
Technique which:
- is used as a cloning alternative
- gets lots of DNA from a little bit
- used for crimes
RT-PCR
Technique which:
-figutes out how much DNA you had to start with versus the control
qPCR
Technique which:
- uses ddA, ddT, ddG, ddC which lack a 3’OH
- terminates growth
- label with fluorescence and separates with elecrophoresis
Sanger Sequence
Technique which:
- chromosome analysis at M phase
- visual DNA - Visualize chromosomal aberrations such as: large deletions, duplications, translocations
FISH
Technique which:
- detects difference between 2 different DNA samples
- detects duplication or deletion of 1-2 Mb
- tests larger chromosome segments/copy
- if equal copy:
- single vs 2 = 0.5:1
- 3 vs 2 = 1.5:1
Comparative Genome Hybridization
Technique which:
- used for examining proteins in cell or tissue extracts
- use proteins with antibody probe
- use fluoro/antibodies to see if proteins are present
Western Blot
This technique is used to identify a gene that has been deleted or if there is a significant deletion/duplication on a particular DNA fragment
Southern Blot
This technique is used to screen a fetus or to confirm a single nucleotide or point mutation or small deletion
ASO
This technique is used to determine the size and abundance of specific RNA in a sample
Northern Blot
This technique is an alternative to cloning that amplifies a DNA, generating copies of a particular DNA sequence
PCR
This technique analyzes small RNA samples; it qualitatively detects gene expression through creation of complementary DNA (cDNA) transcripts from RNA
RT-PCR
This technique is used to amplify and detect or quantify a targeted DNA molecule/quantitatively measure the amplification of DNA using fluorescent probes
q-PCR
This technique is used to sequence any purified DNA using ddNTPs
Sanger Sequencing
This technique is used to visualize chromosomal abberations such as deletions, duplications, and translocations
FISH
This technique is used to examine an entire genome or thousands of genes at one time
Microarray
This technique is used to quickly and efficiently compare two genomic DNA samples arising from two sources, which are most often closely related, because it is suspected that they contain differences in terms of either gains or losses of either whole chromosomes or subchromosomal regions
Comparative Genome Hybridization
This technique is used to analyze the number of transcripts made in a particular cell type, tissue, or disease state relative to another sample
RNA Expression Arrays
This technique is used to examine proteins in cell or tissue extracts to detect how a molecular DNA defect alters the encoded protein to produce the clinical phenotype
Western Blot
This technique uses 24 different chromosome painting probes with different fluorescent dyes that emit at different wavelengths to visualize abnormal chromosomes to diagnose specific chromosomal abberations
Spectral karyotyping
