DNA technology Flashcards
To sequence a piece of DNA with the dideoxy method, you will probably want to use
Fluorescent-labeled dideoxyribonucleotides
You have been instructed by the U.S Department of Health and Human Services to screen the whole population of New York City for the presence of the sickle cell mutation. What method would be best for this project?
Dot blotting
Retroviral vectors are more popular for somatic gene therapy than other viral vectors because
They integrate themselves into the host cell DNA
The steps of classic Southern blotting include (1) denaturation of DNA with alkali, (2) electrophoresis in a cross-linked agarose pr polyacrylamide gel, (3) application of a probe, (4) treatment of DNA with a restriction endonuclease, and (5) blotting of DNA to nitrocellulose paper. The correct sequence of these steps is:
4-2-1-5-3
A child born with multiple congenital malformations. The karyotype appears normal, but the treating physician suspects a deletion or duplication. Which method should be used to screen the genome for copy number variations?
Array comparative genome hybridization
The classic PCR procedure (without probes) can be used to
Diagnose HIV infection in people with risky lifestyles
PCR-based procedures are often preferred over Southern blotting for the prenatal diagnosis of genetic diseases after amniocentesis or chorionic villus sampling. Why?
PCR requires less DNA; therefore, lenghty cell culturing may not be necessary.
In order to genotype a skin color gene in DNA from a 30,000-year0old Neanderthal skeleton, you will definitely have to use
PCR
In order to compare the expression of a large number of genes in rhabdomyosarcoma cells with gene expression in normal skeletal muscle, you have isolated mRNA from the two souces by affinity chromatography on an oligi-dT column. The most direct method for comparing the two mRNA patterns would be
Northern blotting
Linkage studies with closely linked microsatellite polymorphisms are the preferred diagnostic method for the detection of heterozygous carriers of recessive disease genes when
There is allelic heterogeneity, and it is not known which mutation in a known disease gene causes the disease in a family
Alternatively, the mRNAs from the tumor and normal cells can be compared using
cDNA microarrays
An SNP in the coding region of a tumorsuppressor gene was recently discovered. Heterozygous individuals synthesize both the normal and mutant forms of this gene. The normal form codes for a leucine at a particular position within the protein; when the SNP is present, that codon is altered to code for an arginine. Although the SNP was identified at the nucleotide level, you wish to separate and characterize 10mg of each of the proteins. Assuming you had a mixture of the two forms of the protein, what initial approach would you use to separate these two forms?
Ion exchange chromatography
A patient has a disease that leads to hyperexcretion of a protein in the urine. Which methodology is the fastest, easiest, and least expensive to determine the molecular weight of the native protein in the urine
Size exclusion chromatography
A 42-year-old msn presents to the emergency room and receives a neurologic consult for a slowly progressing chorea with some rigidity and seizures. These episodes have occurred in the past, and a deceased mother had choreic neurologic episodes and dementia. You suspect the patient might have Huntington disease, and the hospital has a laboratory equipped to perform RFLP analyses. Which of the following techniques is required to carry out RFLP analysis?
Southern blot
A patient you have been treating for Gaucher disease, a lysosomal storage disease, come into your office wanting to have her relatives tested to determine whether they are carriers of the disorder. The most predominant b-glucosidase mutation, N370S, is expressed in this family. Wanting to do this in the most expeditious and least costly manner would require the use of which one of the following techniques?
Allele-specific oligonucleotide probe hybridization
