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Complementation analysis
Definition - Complementation analysis is used to determine whether two mutations causing the same phenotype are in the same or different genes. If two mutants with the same phenotype produce a wild-type offspring when crossed, the mutations are in different genes (they complement each other).
Addresses:
Are two mutations in the same gene or different genes?
How many genes are involved in a specific biological pathway?
Restriction nucleases
Definition - Restriction nucleases (enzymes) are proteins that cleave DNA at specific sequences into fragments for analysis or cloning.
Addresses:
How can DNA be cut for cloning or sequencing?
What are the specific sequences in a DNA molecule?
SNPs (Single-Nucleotide Polymorphism)
Definition - Single-base differences in the DNA sequence between individuals. They are detected using sequencing or genotyping techniques.
Addresses:
How do genetic variations contribute to diseases or traits?
What are the genetic differences between populations?
Mutations
Definition - Changes in the DNA sequence, can be naturally occurring or induced experimentally to study gene function.
Addresses:
What is the role of a specific gene or protein in a biological process?
How do genetic changes lead to disease?
DNA gel electrophoresis
Definition - Separates DNA fragments by size using an electric field applied to a gel matrix. Smaller fragments migrate faster than larger ones.
Addresses:
What is the size of a DNA fragment?
Has a DNA sample been successfully cut or amplified?
Band/gel shift assays
Definition - Detect protein-DNA interactions: DNA bound to protein migrates more slowly through a gel than free DNA.
Addresses:
Does a protein bind to a specific DNA sequence?
What are the binding affinities of protein-DNA interactions?
Southern Blots
Definition - Detects specific DNA sequences in a sample: DNA is separated by gel electrophoresis, transferred to a membrane, and probed with a labeled DNA sequence.
Addresses:
Is a specific DNA sequence present in a sample?
What is the copy number of a gene?
Northern Blots
Definition - Detects specific RNA sequences: RNA is separated by gel electrophoresis, transferred to a membrane, and probed with a labeled RNA or DNA sequence.
Addresses:
Is a specific RNA sequence expressed in a sample?
How does gene expression vary under different conditions?
Molecular beacons
Definition - Fluorescent probes that emit light when bound to a specific DNA or RNA sequence. They are used for real-time detection of nucleic acids.
Addresses:
Is a specific DNA or RNA sequence present in a sample?
How does gene expression change in real time?
cDNA microarrays
Definition - Measure gene expression levels by hybridizing cDNA from a sample to a chip containing thousands of DNA probes.
Addresses:
Which genes are differentially expressed under specific conditions?
What is the gene expression profile of a tissue or cell type?
Single cell RNA-sequence
Definition - Single cell RNA-Seq measures gene expression in individual cells by sequencing their RNA.
Addresses:
What is the gene expression heterogeneity within a cell population?
How do individual cells differ in their response to stimuli?
Genomics Cloning
Definition - Involves inserting large fragments of genomic DNA into vectors for amplification and study.
Addresses:
What is the function of a specific genomic region?
How can large DNA fragments be studied or manipulated?
cDNA Cloning
Definition - Involves reverse-transcribing RNA into DNA and inserting it into a vector for amplification and study.
Addresses:
What is the sequence of a specific mRNA?
How can a protein-coding gene be expressed in a host organism?
PCR/RT-PCR
Definition:
PCR - Amplifies specific DNA sequences
RT-PCR - Amplifies RNA after reverse transcription into cfDNA
Addresses:
Is a specific DNA or RNA sequence present in a sample?
How does gene expression change under different conditions?
CRISPR-Cas9
Definition -
Addresses:
Used to cure sickle cell disease
