Term
| Which has higher resolution, genetic or physical maps? |
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Definition
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Term
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Definition
| the frequency of recombination between loci |
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Term
| Give two examples of genetic mapping |
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Definition
Restriction Fragment Length Polymorphism
Minisatellites & Microsatellites / Short Tandem Repeats |
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Term
| What are DNA polymorphisms? Why are they useful? |
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Definition
variation in nucleotide sequence between individuals in a population
-can use them as genetic markers in RFLP |
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Term
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Definition
| detecting presence/absence of restriction sites at a locus |
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Term
| What are the two processes involved in RFLP mapping? |
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Definition
-restriction digestion
-Southern hybridization |
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Term
| Describe the steps of RFLP |
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Definition
1. isolate DNA from each ecotype
2. digest DNAs with restriction enzyme
3. separate restriction fragments using electrophoresis
4. transfer the fragments to a nylon membrane
5. hybridize the DNA fragments to a radioactive clone of a gene (Southern blotting)
6. create an autoradiogram by exposing the blot to xrays
7. Compare! |
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Term
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Definition
| highly polymorphic, repetitive DNA with large variability in repeat numbers |
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Term
| After measuring variable number tandem repeats, what can you conclude about the intensity of fluorescence observed at different alleles? |
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Definition
| More intense flouresence means more STR repeats at that locus |
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Term
| What is DNA fingerprinting? |
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Definition
| -using PCR & southern blotting/gel electrophoresis to detect VNTR or STR polymorphisms |
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Term
| What do the individual bands of DNA fingerprints represent? |
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Definition
| different alleles of VNTR/STR loci |
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Term
| When running a paternity test, if a band is common to both parents and observed in the child, can you make a conclusion about the father's relation based on this observation? |
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Definition
| no, because you don't know whether the child inherited the DNA from the mother or father |
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Term
| This physical map is a set of overlapping genomic DNA clones |
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Definition
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Term
| Give 3 methods of ordering genomic clones into contigs |
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Definition
-restriction mapping
-STS mapping
-DNA sequencing |
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Term
True or False
sequence-tagged sites exist in multiple spots throughout the genome |
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Definition
| False, they are unique, each STS is found only in 1 spot! |
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Term
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Definition
| DNA sequences that are mapped both physically and genetically |
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Term
| This type of mapping involves locating genes on chromosomes by in situ hybridization using radioactive probes or fluorescent probes |
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Definition
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Term
| Why don't physical map distances always correlate with genetic distance? |
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Definition
| -frequency of recombination isn't always proportional to molecular distance |
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Term
True or False
Genes are distributed evenly throughout a chromosome |
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Definition
| False, there are gene deserts & gene-dense regions |
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Term
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Definition
single nucleotide polymorphism
-single base pair substitution |
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Term
| Where are most SNP's located? |
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Definition
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Term
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Definition
| a set of SNPs on the same chromosome that are often inherited as a group |
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Term
| What are homologous genes? |
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Definition
| genes derived from a common ancestor |
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Term
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Definition
| homologous genes in different species |
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Term
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Definition
| duplicated gene in one individual which evolves to have a new function |
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Term
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Definition
| the entire set of mRNA transcripts produced by an organism |
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Term
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Definition
| The entire set of proteins encoded by an organism |
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Term
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Definition
1. prepare a microarray with oligonucleotides specific to the genes in question
2. isolate RNA from both control & experimental samples
3. label the samples of cDNA with fluorescent dyes
4. hybridize the probes to the samples
5. observe the different colours of flourescence & compare expression between experimental cells & control cells |
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Term
| Does northern blotting use RNA or DNA? |
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Definition
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Term
| Does southern blotting use RNA or DNA? |
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Definition
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Term
| How might you make use of a protein as a reporter? What is an example of a protein often used a reporter? |
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Definition
-study regulation of gene expression
-substitute an expressed gene with a reporter, which will make it obvious when a gene is turned 'on'
GFP is often used for this! |
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Term
| This type of DNA fragment is synthesized from a messenger RNA template |
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Definition
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Term
| What are the 4 ways expression of eukaryotic DNA is regulated? |
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Definition
-post transcriptionally
-post translationally
-alternative spliceing
-transcriptionally |
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Term
| Where does regulation at a transcriptional level take place? |
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Definition
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Term
| What is a cloning vector? |
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Definition
| A small piece of DNA, usually taken from a virus or plasmid, into which a foreign gene can be inserted for cloning |
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Term
| Why is it important for a cloning vector to have a replication origin? |
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Definition
| So that the foreign gene being inserted can be replicated |
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Term
| In RFLP, _______ are hybridized to ________. |
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Definition
| DNA restriction fragments....radioactive gene clones |
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Term
| Why type of blotting is used in RFLP? |
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Definition
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Term
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Definition
| no! southern blotting is used to visualize the fragments of DNA |
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Term
| Why isn't Southern blotting used when creating a DNA fingerprint from VNTRs or STRs? |
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Definition
| the tandem repeats are amplified using PCR to obtain a measurable amount |
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Term
| What does VNTR stand for? STR? |
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Definition
Variable Number Tandem Repeat
Short Tandem Repeat |
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Term
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Definition
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Term
| What does FISH stand for? It is an example of what kind of mapping? |
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Definition
Flourescent in-situ hybridization
-cytogenic mapping (physical) |
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Term
| In DNA Microarrays, what is labeled and to what is it hybridized? |
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Definition
| cDNA fragments are fluorescently labelled & hybridized to the probes on microarrays |
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Term
| In Microarrays, what are the cDNAs usually derived from? |
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Definition
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Term
| What are some examples of physical mapping? |
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Definition
restriction maps
STS maps
contigs |
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Term
| According to the Human Genome Project, how much of our genomic DNA is transposable? |
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Definition
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Term
| According to the Human Genome Project, how much of our genomic DNA encodes proteins? |
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Definition
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Term
| What is the frequency of SNP between two individuals? |
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Definition
| 1 SNP every 1200 base pairs |
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Term
| VNTRs are known as ______ while STRs are known as _______ |
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Definition
minisatellites
microsatellites |
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Term
| Compare the length of minisatellites to microsatellites |
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Definition
| 10-100 nucleotides vs 2-9 nucleotides |
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Term
| Describe the steps involved with DNA fingerprinting involving VNTRs & SNRs |
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Definition
1. amplify the STRs using flourescently-tagged locus-specific primers (PCR!)
2. Separate the PCR products with gel electrophoresis
3. Compare the PCR products (number of repeats) to the standard alleles of each STR |
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