Biochemistry - Lab Techniques

Question 0

Northern Blot

Answer 0

RNA
RNA is enzymatically cleaved–> electrophoresis and filtered –> radio labeled –> RNA visualized

Indicated the level of mRNA and gene expression

Question 1

Southern Blot

Answer 1

DNA

DNA cleaved into smaller pieces–> electrophoresis and filtered –> DNA is radio labeled –> DNA visualized

Question 2

Western Blot

Answer 2

Protein
Proteins separated and electrophoresis –> labeled with antibody

• used to verify HIV dx after POSITIVE ELISA

Question 3

SouthWestern Blot

Answer 3

identify DNA binding Proteins (transcription factors) using oligonucleotide probes

Question 4

Microarrays

Answer 4

Used to identify gene expression, SNPs, CNV, genetic testing etc

Profiles the DNA by complementary binding with known DNA/RNA sequences

Question 5

ELISA

Answer 5

Detect the presence of an antibody or an antigen
Direct: antibody used to detect presence of an antigen, antibody coupled to a color generating enzyme
Indirect: Antibody or antigen detected, then color generating enzyme used to detect the antigen-antibody complex

Question 6

Karyotyping

Answer 6

Metaphase Chromosomes are detected, stained, and ordered based on various factors
- dx CHROMOSOMAL ABNORMALITIES

Question 7

Floresence in Situ

Answer 7

Complementary binding with DNA/RNA probes to a specific gene of interest
Used for localization of the genes and direct visualization of anomalies

Question 8

Cloning

Answer 8

mRNA –> cDNA via reverse trancscriptase –> inserted into plasmid with antibiotic resistance gene –> antibiotic resistance Bacteria created

Question 9

Cre-lox

Answer 9

Can inducible modify the expression of genes at different points in development

Question 10

RNA interference

Answer 10

dsRNA is synthesized that is complementary to the mRNA of interest. dsRNA is inserted into the human cells, and the dsRNA separates and combines with the host cell mRNA promoting degradation.
- to knock out gene expression