Term
| Nucleic acids can be found in the ___ of an animal cell, while proteins are made on ribosomes associated with the ___, and proteins are modified in the ___. |
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Definition
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Term
| What is the process by which germ cells reproduce, thereby giving us cells with only 1/2 the complement of genetic information? (only 23 chromosomes). |
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Term
| Which of the following IS NOT a base found in RNA? |
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Definition
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Term
Which of the following mRNA sequences was derived from this DNA sequence?
5'-ATGACCGGCAATAAGGGCCGT-3' |
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Definition
| 5'-AUGACCGGCAAUAAGGCCGU-3' |
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Term
| Which of the following represents an accurate series of molecular events in a cell? |
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Definition
| DNA -> mRNA -> tRNA -> protein |
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Term
| Which of the following techniques are used to examine gene expression patterns? |
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Definition
| Northern Blotting and In situ analysis |
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Term
| What is the name of the chemical that intercalates into DNA and is used to visualize DNA by UV light? |
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Definition
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Term
| If you wanted to separate pieces of DNA on an agarose gel and their sizes were: 10,000bp; 15,000bp; and 25,000 kb, what % agarose would you use? |
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Definition
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Term
| Which type of bacterial cell uses heat shock for transformation of plasmid vectors? |
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Definition
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Term
| What type of vector causes lysis of the bacterial cell when it grows within the cell? |
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Definition
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Term
| What type of vector would you use to clone an entire gene including introns and non-coding sequences? |
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Definition
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Term
| What type of library will give you information on mRNA expression in your cell? |
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Definition
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Term
| If you want to make a protein from you cDNA sequence, you must clone your cDNA into what type of vector? |
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Definition
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Term
| What type of vector would you use to clone a piece of your gene that is 5kb, and you would like to find restriction sites within your gene? |
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Definition
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Term
| What are three essential components of a PCR reaction? |
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Definition
| template, primers, and polymerase |
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Term
| Which of the following lists a PCR cycle in its correct order? |
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Definition
| denaturation, annealing, extension |
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Term
| Which primer has the highest Tm? |
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Definition
5'-GCTGTGCGTAGGATGCGTATG-3'
12(4)=48 + 18=66
4(G+C) + 2(A+T) |
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Term
| Which of the following are non-covalent bonds? |
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Definition
| hydrogen bonds, ionic bonds, and van der waals |
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Term
| Which of the following is NOT a function of chromosomes? |
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Definition
| Provide a template for translation machinery |
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Term
| Which of the following are true about histones? |
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Definition
-Exist as octamers when bound to DNA
-Histone proteins are recycled during DNA replication and reassembled once the DNA strand is replicated
-Acetylation of histones allows DNA to be more loosely packed allowing for transcription of genes |
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Term
| If you isolate cells and expose them to a low concentration of DNAse I, then you separate DNA from bound protein, and run that DNA out on a gel, what will you see? |
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Definition
| Genes that were not able to be transcribed |
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Term
| What is the process by which all our cells reproduce to generate an exact duplicate cell? |
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Definition
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Term
| What of the following IS NOT a base found in DNA? |
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Definition
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Term
| If you wanted to separate pieces of DNA on an agarose gel and their sizes were: 500bp, 890bp, and 1000bp, what % agarose would you use? |
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Definition
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Term
| Which statement is true regarding phage vectors? |
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Definition
-They kill bacteria as they replicate
-They can hold very large pieces of DNA |
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Term
| Which is not an essential component of a PCR reaction? |
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Definition
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Term
| Which of the following are covalent bonds? |
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Definition
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Term
| Three things you need in a vector to make it a good tool for molecular studies. |
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Definition
-Origin of replication (ori) appropriate for host cell
-Selectable marker allowing for isolation of cells carrying the vector
-mechanism allowing segregation into daughter cells |
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Term
| 2 DNA modifying enzymes that you would use when cloning your DNA into a vector prior to transforming that recominant DNA into bacteria. |
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Definition
-Restriction enzymes: digest target DNA and digest vector w/ same enzyme needed to clone pieces of DNA into vector.
-CIAP: removes 5' phosphate to precent self-annealing (closing up on itself) |
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Term
| When running DNA on a gel, what type of charge attracts the DNA? What property of DNA allows this to happen? |
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Definition
-DNA is negatively charged, so attracted to cathode (positive) end of the gel and runs towards it.
-Property that allows this is the phosphate backbone on DNA |
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Term
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Definition
Detects DNA using solid membrane support
1) Cut DNA
2)separate DNA fragments on gel
3)detature d.s DNA into gel
4)transfer DNA to solid support (nylon membrane)
5)hybridize with labeled DNA squence (probe)
6)wash blot
7)detection |
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Term
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Definition
| Similar to southern blotting, but transfers RNA. Use DNA prob against total RNA to determine if a gene is expressed (made into cDNA) and what size transcript (no introns) |
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Term
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Definition
| Transfer proteins into solid support, mediated by a current. Usually are using antibodies to recognize proteins. |
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Term
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Definition
| RNA probe hybridizing to mRNA in a cell |
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Term
| FISH (fluorescence in situ hybridization |
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Definition
| localize where RNA is specifically |
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Term
| WISH (whole mount in situ hybridization |
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Definition
localize where genes are being expressed.
overall picture of regions of expression; sectioning the whole mount provides added detail |
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Term
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Definition
| tells exactly if DNA sequence protected or bound by a protein |
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Term
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Definition
Reverse transcriptase PCR
-Use total RNA, make cDNA (reverse transcribe) and amplify cDNA |
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Term
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Definition
real-time PCR
Quantitates the initial amount of the template. Used with reverse transcriptase to determine amount of message in a sample. |
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Term
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Definition
| use antibodies to detect proteins; detect exactly where protein is being expressed |
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Term
| Maxam0Gilbert method DNA sequencing |
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Definition
| DNA sequence end-labeled with phosphate and chemically cleaved to leave signature pattern of bands (not commonly used) |
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Term
| Sanger method DNA sequencing |
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Definition
| DNA sequence annealed to oligonucleotide primer, then extended by DNA polymerase using mix of dNTP and ddNTP (chain terminating) substrates |
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Term
| Why Maxim gillbert not commonly used? |
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Definition
Fragments need to be isolated, labeled, cleaved, and purified: tedious!
Used to determine where proteins bind to DNA
Nucleotides protected from chemical cleavage bc under protein, so not available to run on gel. |
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Term
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Definition
Restriction fragment length polymorphism
rough matches to identify samples |
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Term
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Definition
primer extension reaction to amplify specific nucleic acid sequences
-make many copies of DNA
-both strands are amplified/copied..sso EXPONENTIAL increase in # copies of gene |
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Term
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Definition
obtain large amount of material for sequencing
obtain large amount of material for cloning
detect very small amounts of dna in sample
-forensics
-disease ID
-transgenic analysis |
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Term
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Definition
| try to clone gene encoding a protein |
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Term
| Radioactive nucleotide labeling |
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Definition
common
-health risk, longer exposure time for detection, half life-unstable (decays) |
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Term
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Definition
| make hundreds of copies of dna, detects on radiographic film |
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Term
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Definition
non-radioactive labeling
-can make antibodies to it and be coupled to nucleotides and incorporated into nucleic acids
-safe, no decay, short exposure time, detection by color rxns and chemiluminescence |
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Term
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Definition
10-20kb
larger plasmids, large insert- but harder to get into the cell (transformation) and prone to bacterial rearrangements
-various antibiotic resistance genes |
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Term
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Definition
20-25kb
bacteriophage
lytic, kill host after replicate dna
-package very large inserts
-require certain size of insert DNA to package (control for big pieces only) |
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Term
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Definition
45 kb
plasmid with cos sites
-cos sites allow for a bigger insert than reg. plasmid |
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Term
| BAC-bacterial artifical chromosome vector |
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Definition
hold much more dna than plasmid or viral vector
-up to 90 kb of gDNA |
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Term
| YAC- yeast artificial chromosome vector |
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Definition
100's of kb
ARE chromosomes
difficult to maipulare and introduce into yeast w/high efficiency
-gDNA libraries |
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Term
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Definition
| catalyses liigation of annealed DNA duplexes |
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Term
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Definition
| removes phosphate groups. helps self annealing |
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Term
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Definition
| phosphorolates (adds phosphorous) |
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Term
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Definition
| copy nucleic acid chains into daughter strand using template |
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Term
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Definition
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Definition
| make DNA copy. DNA replication |
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