Biochem lab techniques Flashcards
(14 cards)
Blotting procedures
SNoW DRoP
Southern = DNA
Northern = RNA
Western= protein
PCR steps
Amplify desired DNA fragment
- Denaturation
- Annealing
- Elongation
Flow cytometry commonly used in?
Hematologic abnormalities and immunodeficiencies
Flow cytometry how work?
- Tag cells with antibody specific to surface or intracellular protein
- Antibody tag with fluorescent dye
- Histogram or scatter plot shows light scatter and intensity of fluorescence
Microarray detect what?
SNP (single nucleotide polymorphism) and CNVs (copy number variations)
Microarray used for?
Genotyping, clinical genetic tests, forensic analysis, cancer mututions, and genetic linkage analysis
ELISA tests for?
- Presence of specific antigen (like HBsAg) or antibody (like anti-HBs) in blood
- color change= positive test
Karyotyping
Metaphase chromosomes looked at
Karyotyping used for
Chromosomal imbalances (autosomal trisomies, sex chromosome disorders)
FISH used for what kind of diagnoses?
Chromosomal anomalies at molecular level
- Microdeletion= no fluorescence compared to same locus on second copy
- Translocation= fluorescence outside the original chromosome
- Duplication= extra site of fluorescence on one chromosome
Cloning method steps
- Isolate mRNA
- Expose to reverse transcriptase make cDNA
- Insert into antibiotic resistant bacterial plasmids
- Transform plasmid to bacteria
- Surviving bacteria on antibiotic medium produce cloned DNA
Gene expression modification strategies
Homologous recombination
Knock-out= removing gene, taking it out
Knock-in= inserting a gene
RNA interference does what?
Knock down gene expression by promoting degradation of target mRNA
Cre-lox system
Manipulate genes at specific developmental points
