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Diagnostic Tests
WVSOM -- Biochem -- Diagnostic Tests
| Question | Answer |
|---|---|
| PCR | Polymerase chain reaction. To amplify a piece of DNA by in vitro enzymatic replication. |
| Reducing sugar test | Fehling's Test. Does not reduce sucrose! |
| Southern Blot | check for the presence of a DNA sequence in a DNA sample. Utilizes gel electrophoresis and after the DNA is put thru the electrophoresis a probe is inserted and the DNA sequence appears. |
| Northern Blot | Like Southern Blot except Measures for RNA |
| Western Blot | Measures Proteins. Like Southern Blot except used to meaure proteins. Antibodies used as the probe. |
| Probe | a single-stranded piece of DNA or RNA that is used to visualize complementary sequences within DNA. |
| Fredrick Sanger Method | dideoxynucleotides triphosphates (ddNTPs) as DNA chain terminators. Dideoxynucleotides lack a 3’and a 2’ hydroxyl group which halts polymerization. Used to halt the reaction in polymerization. Sequences DNA |
| How is PCR used to diagnose disease? | Allele-Specific Oligonucleotide Probes. Detection of polymorphisms caused by repetitive DNA. Testing for mutations by PCR. DNA Chips |
| Why is PCR important? | Ability to amplify and see mutations in DNA. |
| How much does PCR amplify DNA? | DNA is amplified over a million fold (1,000,000). Restriction fragment length polymorphisms. |
| RFLP | RFLP analysis is a technique that can identify some differences in sequence. Used in maternity testing and forensics. Sickle Cell can also be detected this way. |
| DNA CHIP Technology | Ability to place DNA on a chip where up to 10,000 copies can be stored. Each gene then can be tested to find what mutations are on each gene. |
| Gene Knockout | Using mice there is a way to find out what genes do. We knock out gene of interest and see what happens. |
| Anophthalmia/microphthalmia | loss or severe reduction of orbital structures (~1/10,000) |
| Karyotypes | spin down DNA and isolate from WBC and treat with mitosis to see a chromosomal display |
| FISH | florescent marker to a probe and find an abnormality on the chromosome |
| RNAi | targets proteins that are overexpressed and it binds to that protein and decreases the protein expression. |
| What can be treated with RNAi? | HIV Huntington's Disease Pandemic Flu Liver Cancers Cancer Respiratory Syncytial Virus |
| recombinant DNA | artificial DNA |
| transgenic | genetically modified |
| Why is genetic susceptibility used? | To see what diseases a person may get thru genetics |
| Epigenetics | Heritable changes in gene expression caused by mechanisms other than changes in the DNA sequence |
| What are 2 examples of epigenetics? | Histone modifications and DNA methylzation |
| What is a current epigentic test used for? | prostrate cancer |
| What is gene therapy? | Correct genetic disorders by introducing a normal functioning copy of the defective gene |
| somatic | refers to cells of the body |
| In vivo into human somatic cells? | Delivery method must be targeted to specific tissue. Methods include injection or gene gun, viral vectors. “Naked” DNA is very quickly chopped up in the body so vehicles are used (liposomes, nanoparticles, etc) |
| Ex vivo introduction of genes into a human somatic cells | Ex vivo Cells are removed from a patient and the gene is introduced Methods include liposomes, electroporation, gene guns, viral vectors, nanoparticals, etc Cells are then transplanted back into the patient |
| Why use viral vectors? | Viruses inject their genetic material into the host cell as part of their life cycle and we can alter dna/rna of viruses |
| What are retroviral vectors? | Gene is integrated into the host genome so daughter cells will have it |
| What is the problem with retroviral vectors? | Risk of insertional mutagenesis |
| What is being used besides genes with retroviral vectors? | transposons |
| What are problems with Gene Therapy? | The DNA needs to stay around and be transcribed a long time in order to be effective Immune responses can occur Potential for reactivation of viruses It’s hard to control DNA! Disorders caused by multiple genetic changes are more difficult to treat us |
| What are stem cells? | Undifferentiated cells with the capacity for self-renewal and the ability to develop into differentiated cells |
| Totipotent | Cells produced by the first few divisions of a fertilized egg. Can become any cell type including extraembyronic (such as placenta) |
| Pluripotent | Isolated from the inner cell mass of a blastocyst Can become any cell type except extraembryonic cells |
| Multipotent | Can give rise to a limited number of cell types, generally in the same family (such as hematopoietic stem cells) Found in more developed fetuses and adults |
| Unipotent | Can become only one cell type (such as muscle stem cells |
| Somatic cell nuclear transfer | cloning |
| What is a common stem cell therapy use? | bone marrow |
| Autologous | patient transplanted with their own |
| Allogenic | Patient transplanted with stem cells from a matched donor |
| Umbilicord blood | another possible source of embryonic like stem cells |
Created by:
tjamrose