Term
| What does DNA sequencing do? |
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Definition
| Determines the exact nucleotide composition of a DNA fragment |
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Term
| What are the 4 applications of DNA sequencing? |
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Definition
Diagnostic testing
Forensics
Research
Biotechnology to produce therapeutics |
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Term
| What are the Research applications of DNA sequencing? |
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Definition
Human Genome Project
Understanding genetics, diseases
Allow determination of amino acid sequence which affects protein function |
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Term
| What are the 4 essential components of DNA synthesis in vitro? |
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Definition
*Template DNA
*DNA polymerase
*DNA primer complementary to template DNA
*All 4 deoxynucleotide triphosphates |
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Term
| List the Process of DNA synthesis in Vitro |
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Definition
1. DNA polymerase synthesizes new DNA from template
2. DNA polymerase requires an existing primer; cannot initiate DNA synthesis without primer
3. DNA synthesis occurs only in 5' to 3' direction
4. Polymerase reaction requires all 4 dNTPs |
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Term
| Dideox nucleotide (ddNTPs) have ____ present at the 3' carbon of the pentose sugar instead of a OH group |
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Definition
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Term
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Definition
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Term
True or False:
ddNTPs are added to polymer in the same manner as dNTPs |
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Definition
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Term
True or False:
ddNTPs cannot form bond with next nucleotide |
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Definition
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Term
True or False:
dNTPs cannot form bond with next nucleotide |
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Definition
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Term
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Definition
| Terminate DNA chain elongation |
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Term
| What are the 4 Components Required for the Dideoxy sequencing reaction? |
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Definition
1. Template DNA
2. DNA primers complementary to template DNA
3. All 4 normal dNTPs
4. DNA polymerase |
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Term
What fluorescent color is associated with the following:
ddATP |
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Definition
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Term
What fluorescent color is associated with the following:
ddTTP |
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Definition
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Term
What fluorescent color is associated with the following:
ddGTP |
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Definition
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Term
What fluorescent color is associated with the following:
ddCTP |
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Definition
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Term
True or False:
When dideoxy sequencing systems have 4 reaction tubes everything is the same in each tube except only one fluorescent ddNTP/tube |
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Definition
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Term
True or False:
Some Dideoxy sequencing systems have all 4 fluorescent ddNTPs in the same reaction tube |
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Definition
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Term
| 4 Step Process of DNA dideoxy sequencing (chain termination method) |
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Definition
1. Chain elongation proceeds normally as long as polymerase uses dNTPs
2. Periodically a ddNTP is incorporated into the polymer & chain elongation stops
3. DNA strands vary in nucleotide length
4. After incubation period, fragments sorted by size using electrophoresis |
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Term
True or False:
PAGE is the traditional method with the shortest DNA fragment |
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Definition
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Term
| What does PAGE DNA fragment contain? |
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Definition
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Term
True or False:
PAGE shortest DNA fragment moves fast |
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Definition
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Term
| How does the Longest (and largest) DNA fragments move? |
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Definition
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Term
True or False:
Nucleotide at 5' end of a strand can be determined by the fluorescent dye attached to the ddNTP |
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Definition
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Term
True or False:
Nucleotide at 3' end of a strand can be determined by the fluorescent dye attached to the ddNTP |
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Definition
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Term
| For an Automated DNA sequencer, can you use capillary electrophoresis |
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Definition
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Term
| Are all 4 ddNTPs in same reaction tube with automated DNA sequencers? |
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Definition
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Term
| Automated DNA sequencers - Fragments are separated by size as they pass through what? |
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Definition
| A very long special capillary tube |
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Term
| Automated DNA sequencers - At the end of the capillary tube _______ is read and recorded as each fragment passes through detector |
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Definition
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Term
| Automated DNA sequencers - What analyzes the data and reports DNA sequence? |
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Definition
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Term
| Clinical application - HIV Genotyping what is DNA Sequencing used to determine? |
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Definition
| Resistance to anti-retroviral drugs |
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Term
| Clinical application - HIV Genotyping what is the specimen? |
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Definition
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Term
| Clinical application - HIV Genotyping what can you use to make complementary DNA since HIV is a RNA virus? |
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Definition
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Term
| Clinical application - HIV Genotyping what do you sequence? |
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Definition
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Term
| Clinical application - HIV Genotyping When you analyze the data for mutations what are you trying to see if they are associated with? |
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Definition
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