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18 Cards in this Set
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- Back
- 3rd side (hint)
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Molecular diagnosis of feline B-cell neoplasia:
How can PCR be used to distinguish a neoplastic from a nonneoplastic cell population? Possible PCR / PAGE results and how to interpret them? What is the importance of duplicate reactions for each sample? |
This article: assessment of diversity within complementarity determining region 3 (CDR3) of the IGH V gene of B cells.
PCR / PAGE: 1 reproducible distinct band = clonal = neoplastic. 2 bands = both alleles clonally rearranged = neoplastic. Smear = polyclonal. 3-5 distinct and reproducible bands = oligoclonality; restricted clonal diversity; typically associated with some inflammatory conditions, but also with immunosuppressive and premalignant states. One or more distinct but nonreproducible bands = pseudoclonality; random amplification of limiting target B-cell DNA. In the absence of duplicate reactions for each sample, pseudoclonality can result in false-positive monoclonal results. |
Vet Pathol 42:5, 2005
see PAGE figures! |
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What is single strand conformation polymorphism and what is its use?
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Mutation screening technique.
Thermally denatured DNA is electrophoresed, and those single stranded DNAs that take up an altered conformation show up as aberrantly migrating bands on the electrophoretic gel. |
J Clin Pathol:Mol Pathol 2002:55:73-79
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What is DNA heteroduplex analysis?
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Technique which exploits the difference in electrophoretic mobility between double stranded DNAs composed of two normal sequences, compared with those composed of one normal and one abnormal sequence (heteroduplex) to detect mutations.
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J Clin Pathol:Mol Pathol 2002;55:73-79
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Which 2 techniques detect mutations by exploiting the different denaturation profile of mutant DNA to its normal counterpart?
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Denaturing gradient gel electrophoresis (DGGE) and thermal gradient gel electrophoresis (TGGE).
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J Clin Pathol:Mol Pathol 2002;55:73-79
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What is the amplification refractory mutation system (ARMS)?
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Mutation detection technique where the PCR oligonucleotide primers are designed to be specific for one mutant sequence variant or another, so that the PCR reaction will specifically detect one or the other variant.
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J Clin Pathol:Mol Pathol 2002;55:73-79
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Which technique involving a special endonuclease can be used to detect abnormal cytosine methylation?
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Methylation sensitive restriction endonuclease. If methylated -> no digestion -> PCR amplification across methylation site allowed.
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J Clin Pathol:Mol Pathol 2002;55:73-79
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Role of epigenetic events in cancer:
methylation status of cancer cell DNA vs normal cells? |
Normal cells: CpG islands hypomethylated; CpG methylated.
Cancerous cells: CpG islands hypermethylated, CpG unmethylated. |
Nature Reviews Genetics Vol 3 june 2002 p.415
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What is the Shtrasburg method?
What is it used for? Did not work in which animal species? |
Polyacrylamide gel electrophoresis (PAGE) on formalin-fixed paraffin-embedded tissue extracts. Bands consistent with species-specific AA.
To identify AA and distinguish it from other types of amyloid proteins. Did not work in mice, hamsters, guinea pig. (Worked in cheetah, cat, cow, duck, dog, goose, chicken, turaco). |
Vet Pathol 42:132-139 (2005)
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Terminology: mice in which a gene has been added?
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Transgenic mice.
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Summer School 2006 Cantor
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What are knockout/knockin mice?
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Mice in which the mouse gene has been deleted and replaced by the human gene (sometimes called "humanized mice")
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Summer School 2006 Cantor
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Generation interval of lab mice?
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3 months
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Summer School 2006 Cantor
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What is Cre?
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A DNA recombinase that recognizes specific DNA sequences (LoxP); it puts together pieces of DNA.
Used in conditional knockout; Cre is under control of specific promoters. May be inducible by e.g. tetracycline, or tissue-specific e.g. cardiac-specific alpha-MHC. |
Cantor Summer School 2006
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Describe a Cre-Lox breeding plan.
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Promotor-Cre mice x Floxed gene mice.
Example: tc-Cre x Floxed insulin receptor. tc = tetracyclin; tc-inducible KO. During any stage of life. |
Cantor Summer School 2006
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What is "top shelf syndrome"?
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Retinal damage in mice kept in top cages, closer to the ceiling lights. More common in albino mice such asl BALB/c.
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Cantor Summer School 2006
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What is the significance of Pneumocystis carinii pulmonary infection in knockout mice?
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It suggests immunocompromise
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Cantor Summer School 2006
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What are ribozymes?
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Segments of RNA that act as enzymes and cleave mRNA.
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Cantor Summer School 2006
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What is insertional mutagenesis?
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The random (not targeted!) insertion of a mutated gene into the host genome.
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Cantor Summer School 2006
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What is genetic drift?
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The accumulation of random mutations after many generations.
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Cantor Summer School 2006
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