Term
| how do we transfer fragments of dna into nitrocellulose? |
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Definition
electrophorese
use filter paper and salt water |
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Term
| what makes a probe high stringency? |
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Definition
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|
Term
| what allows for the detection of a radioactively labeled probe? |
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Definition
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Term
| how many cycles creates a million copies in pcr |
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Definition
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Term
| what is the latest discovery in regards to pcr? |
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Definition
| a polymerase that doesn't denature at the high temps needed to do pcr. this is useful because we need a 93 temp to denature the dna but this also denatures the average polymerase. because of this geneticists used to have to add more polymerase after each denature step. |
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Term
| what is the other name for western blotting? |
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Definition
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Term
| what are the three ways to locate a specific sequence in a genome? |
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Definition
nucleic acid hybridization
western blotting
prc |
|
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Term
| what are the three types of nucleic acid hybridization? |
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Definition
southern hybridization (DNA)
colony/plaque hybridization (DNA)
northern hybridization (mRNA) |
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Term
| what is the simplest way to create a BASIC restriction map? |
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Definition
| digest a genome with a particular restriction endonuclease and then electrophorese it |
|
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Term
| what is the most useful technique for creating a restriction map? |
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Definition
| partially digest a genome with a specific restriction endonuclease. this will allow for all the possible fragments to show up and then we can deduce their order |
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Term
| what does labeling do for creating a restriction map? |
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Definition
| labeling with p32 allows us to see which fragments are on the end of a specific piece of dna |
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Term
| explain a double digest (the process) |
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Definition
1. complete digest with one restriction endonuclease
2. partially digest with the same restriction endonuclease
3. complete digest with different restriction endonuclease
4. completely digest with both restriction endonucleases |
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Term
|
Definition
| restriction fragment length polymorphism |
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Term
|
Definition
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Term
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Definition
| when a short sequence is repeated a variable number of times |
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Term
|
Definition
variable number of tandem repeats
minisatellite |
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|
Term
| how big are microsatellites, VNTR? |
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Definition
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Term
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Definition
| missing or having an extra restriction site |
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Term
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Definition
| pattern of VNTR (minisatellites) at different loci |
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Term
| What is the most common method for DNA sequencing? |
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Definition
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Term
| What are the steps to sequence the DNA of something using the Dideoxynucleotide Method |
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Definition
1. separate DNA into four vials
2. each vial has all 4 of the deoxyribonucleotides
3. each individual vial has one of the for didoexyribonucleotides
4. in each test tube a different length fragment occurs
5. Electrophose all the fragments to get a sequencing ladder gel
6. based on how long the fragment is and what dideoxyribonucleotide was in that vial we can deduce the relative order of each base |
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Term
| What are some innovations to DNA Sequencing? |
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Definition
1. Using plasmids: insert DNA you want to sequence, the plasmid already has an oriC and just throw in a primer
2. PCR: amplify gene, separate into four vials, add on DDoxy into each, add primer
3. Dye: attach different flourescent dye to each DDoxy, then you can do it all in one test tube
4. Hook up to a computer |
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