Term
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Definition
| this is used to amplify a desired fragment of DNA in 3 steps. 1) denaturation - DNA is denatured by heating to generate 2 separate strands. 2) annealing - during cooling, excess premade DNA primers anneal to a specific sequence on each strand to be amplified. 3) elongation - heat-stable DNA polymerase replicates the DNA sequence following each primer. this is then replicated multiple times for sequence amplification. agarose gel electrophoresis is used for size separation of PCR products and compared against a DNA ladder. |
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Definition
| DNA is electrophoresed, then transferred to a filter. radiolabeled DNA probes are then added and detected. |
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Term
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Definition
| same as southern blot, but for RNA |
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Term
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Definition
| same as a southern blot but for protein. SNoW DRoP = southern, northern, western, DNA, RNA, Protein. |
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Term
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Definition
| thousands of nucleic acid sequences are arranged in grids/glass/silicon. DNA/RNA probes are hybridized to the chip. this is used to profile gene expression levels of thousands of genes simultaneously to study certain diseases and treatments. this is able to detect single nucleotide polymorphisms (SNPs) for many conditions. |
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Term
| what is an enzyme linked immunosorbent assay? |
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Definition
| ELISA tests are rapid immunologic tests for antigen-antibody reactivity. the pts blood sample is probed w/either a test antigen or test antibody and a positive interaction will give an intense color. |
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Term
| what is fluorescence in situ hybridization? |
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Definition
| a fluorescent DNA/RNA probe binds to a specific site of interest. this is used for specific localization of genes and direct visualization of anomalies at a molecular level (when deletion is too small to be visualized by karyotype). if there is no fluorescence - the gene has been deleted. |
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Term
| what are 3 cloning methods? |
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Definition
| 1) DNA fragments are inserted into bacterial plasmids which contain antibiotic resistance genes - these plasmids can then be selected for by using the antibiotic and amplified. 2) restriction enzymes cleave the DNA at 4- to 6-bp palindromic sequences, allowing for insertion of a fragment into a plasmid. 3) tissue mRNA is isolated and exposed to reverse transcriptase, forming a cDNA (lacks introns) library. |
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Term
| what is constitutive transgenic gene expression modification? |
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Definition
| random insertion of the gene into a mouse genome |
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Term
| what is conditional transgenic gene expression modification? |
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Definition
| targeted insertion or deletion of a gene through homologous recombination w/mouse gene |
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Term
| what is the cre-lox system (type of transgenic gene expression modification)? |
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Definition
| this can inducibly manipulate genes at specific developmental points using an antibiotic-controlled promoter |
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Term
| what is RNAi (type of transgenic gene expression modification)? |
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Definition
| dsRNA which is synthesized complementary to the mRNA sequence of interest. when transfected into human cells, dsRNA separates and promotes degradation of target mRNA - knocking down gene expression |
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