Applications of Molecular Genetics Only

Chromosomal Karyotyping

Chromosomal Karyotyping

A cytogenetic technique used to analyze the number and morphology of nuclear chromosomes in any cell type.

Metaphase Chromosomes

In the phase of the cell cycle, we can group chromosomes by size

"p" arm and "q" arm

The two arms of a chromosome

Aneuploidy causes this

Gross/large structural changes of the entire genome

Philadelphia chromosome

A specific anomaly found in chronic myeloid leukemia (CML) that can be diagnosed using chromosomal karyotyping.

"G"-Banded Karyotyping

A technique that uses Giemsa staining to create a banding pattern on chromosomes

Heterochromatin

Darkly stained regions of chromosomes that are gene-poor

Euchromatin

Lightly stained regions of chromosomes that are gene-rich

Fluorescence In Situ Hybridization (FISH)

A fluorescence-based technique that allows for the detection and localization of specific DNA sequences on chromosomes.

Denatured labeled fluorescent probe

A DNA probe complementary to the gene of interest that is added to the chromosomal preparation in FISH.

Fluorescent antibody

Binds to the DNA probe in FISH and is labeled with a fluorescent dye.

Counterstaining

Staining chromosomes with a fluorescent dye of a different color to visualize regions of interest in FISH.

Interphase FISH

Performed on non-dividing cells

Spectral Karyotyping (SKY) or Multiplex-FISH (M-FISH)

A technique that uses various combinations of fluorescent dyes to label individual chromosomes

Array-based Comparative Genome Hybridization (aCGH)

A technique that compares the patient's genome to a control reference to assess copy number variations and structural abnormalities.

Whole-Genome Sequencing

Analyzing millions of sequence reads and aligning them to the reference genome to diagnose aneuploidy or structural chromosome abnormalities.

Molecular cloning

A technique that creates a DNA clone from multiple DNA sources and amplifies it in a living system.

Restriction Endonuclease Digestion

Cleaving dsDNA using specific enzymes

Cloning Plasmid

A small circular dsDNA molecule that replicates independently of the host chromosome and contains essential elements for cloning.

Muenke syndrome

A craniosynostosis syndrome caused by a single mutation in the FGFR3 gene

Complementary DNA (cDNA) Library

A library of DNA molecules that are reverse transcribed from mRNA

Polymerase Chain Reaction (PCR)

A technique used to exponentially amplify DNA through cycling

SNP

Single nucleotide differences that fit the definition of polymorphism

dPCR

Digital PCR

Gel Electrophoresis

A technique used to separate charged samples of interest according to size.

DNA Microarrays

Used to analyze the production of thousands of different mRNAs in a single experiment

Flow Cytometry

Cells are tagged with antibodies specific to surface or intracellular proteins

Leukemia

A type of cancer that affects the blood and bone marrow

ELISA

Enzyme-linked immunosorbent assay

Southern Blotting

A technique used to detect specific DNA sequences by separating DNA fragments based on size and transferring them onto a membrane for analysis.

Western Blot

A technique used to detect specific proteins in a sample using antibodies.

DNA Sequencing

The process of determining the precise order of nucleotides in a DNA molecule.

Next-Generation Sequencing

Second-generation sequencing methods that rely on massively parallel sequencing reactions

Diagnosis

The identification of a disease or condition based on signs

Cancer Screening

The process of testing individuals for cancer before symptoms appear

CRISPR/Cas9

A genome editing tool derived from bacteria that consists of a guide RNA (gRNA) and an endonuclease (Cas9) used to make targeted changes to DNA sequences.

Nonhomologous end joining (NHEJ)

A repair mechanism that fixes imperfectly cut DNA segments by joining the broken ends together

Homology-directed repair (HDR)

A repair mechanism that uses a donor DNA sequence to fill in gaps created by DNA cleavage

Antibody

A protein produced by the immune system that can recognize and bind to specific target molecules

Enzyme-linked immunosorbent assay (ELISA)

An immunologic test used to detect the presence of specific antigens or antibodies in a patient's blood sample by using an antibody linked to an enzyme.

Transgenic

Referring to an organism that has had foreign DNA inserted into its genome.

Knock-out

The removal of a specific gene from an organism's genome.

Knock-in

The insertion of a specific gene into an organism's genome.

RNA interference (RNAi)

A process in which small RNA molecules target and inhibit the expression of specific genes.

What method could be used to determine a small genomic section when sample size is low?

Use of Sanger Sequencing

dPCR

enables the most accurate, precise and sensitive detection of specific nucleic acid sequences due to its partitioning effect. Reaction partitioning allows an effective increase of rare target abundance by diluting the background (wild-type) molecules.