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B1 Etc.
extra stuff that didn't fit in cases
| Question | Answer |
|---|---|
| What is cytogenetic analysis? | Involves making a KARYOTYPE showing chromosomal makeup of an individual somatic cell, inc. CHROMOSOMAL NUMBER, ARRANGEMENT, SIZE & STRUCTURE. |
| In what phase of the cell cycle are cells during cytogenetic analysis? | METAPHASE - b/c it's condensed & easier to see. |
| What stain is used to visualize chromosomal bands in cytogenetic analysis? | MGG - different chromosomes naturally stain differently, which allows for visual pairing. |
| What might a mutation look like in cytogenetic analysis? | Discrepancy btn size & banding patterns. |
| What does a normal karyotype look like on cytogenetic analysis? | 22 pairs autosomal chromosomes & 1 pair sex chromosomes |
| What are the advantages of cytogenetic analysis? | fairly simple, been used for years |
| What are the disadvantages of cytogenetic analysis? | time intensive, not as sensitive as newer methods, can’t detect specific BCR-ABL products |
| What does FISH stand for? | Fluorescence In-Situ Hybridization |
| What is FISH used for? | to ID specific parts/mutations on a chromosome, & to find the location of a gene & determine if translocation occurred |
| What is commonly used first to find the presence of Ph+ chromosome? | FISH |
| In what phase of the cell cycle should cells be in FISH? | METAPHASE so that chromosomes don’t overlap |
| What are the advantages of FISH? | quick, commercially available kits, sensitive |
| What are the disadvantages of FISH? | can’t specify different mutations |
| What does PCR stand for? | Polymerase Chain Reaction |
| What is PCR used for? | AMPLIFY DNA sequences. Requires only a sm amt of DNA. Quickly becoming most widely used method of BCR-ABL Dx. |
| What are the advantages of PCR? | quick, primers commercially available, can detect specific mutations |
| What are some specific advantages of PCR? | Can detect much smaller mutations than cytogenic analysis. Can find one BCR/ABL chromosome in a sample of 500,000 cells!!! |
| What is RT-PCR? | Reverse Transcriptase PCR |
| What is RT-PCR used for? | Can see if mRNA is being produced; this references proteins being used. |
| How does RT-PCR work? | Use RT to convert mRNA to DNA then amplify |
| What is the advantage of RT-PCR? | Only see EXONS – so better idea of what PTN will be |
| What is Southern Blot? | detects DNA on gel w/ fluorescently labeled probes |
| What is Northern Blot? | detects RNA on gel w/ fluorescently labeled probes |
| What is Western Blot? | detects protein on gel w/ fluorescently labeled probes |
| How do you do a blot? | Slide product in gel & blot – time intensive so not used very much. Not all diagnostic labs have the equipment |
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