Term
| What does reverse transcriptase do? |
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Definition
| make DNA from RNA, DNA polymerase looking for retrovirus |
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Term
| Number of targets needed for a single PCR target? |
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Definition
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Term
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Definition
| steam vents in yellowstone |
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Term
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Definition
| oligonucleotides to start chain rxn |
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Term
| optimal temp for taq polymerase |
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Definition
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Term
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Definition
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Term
| temp for primer annealing |
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Definition
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Term
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Definition
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Term
| length of time for one cycle |
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Definition
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Term
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Definition
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Term
| long or short target more efficient? |
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Definition
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Term
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Definition
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Term
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Definition
| what youre trying to copy |
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Term
| how many sets of long template? |
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Definition
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Term
| how many sets of intermediate template? |
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Definition
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Term
| what is a precise length template (amplicon)? |
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Definition
| much more common than long and intermediate, what you're looking for, PCR product |
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Term
| PCR work is unidirectional. what order? rules? |
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Definition
reagent to extraction to PCR
-one tube open at a time
-keep tops on as much as possible
-reagents in small aliquots, use once
-clean with bleach
-use disposable plastic ware
-pulse spin tube
-avoid splashing and aerisols |
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Term
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Definition
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Term
| What is complimentary DNA and how is it different from DNA? |
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Definition
| cDNA-DNA made from RNA, lacks introns |
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Term
| Applications for reverse transcriptase |
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Definition
| looking for HIV, HCV, and gene expressions in the disease process..(?) |
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Term
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Definition
poly (T) for gene expression
Viral primer:specific |
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Term
| Multiplex PCR-number of targets and primers |
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Definition
| 2+ targets, primer for each target, IDs species and detects target |
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Term
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Definition
| quantitative, can see test and it's happening |
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Term
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Definition
| semi-quantitative, amplicon detected at end of rxn, need a post test medium like electrophoresis |
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Term
| what are the stages of PCR |
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Definition
exponential- precise specific doubling
linear-variable, slower
plateau- end point, rxn stop |
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Term
| What is cycle threshold (Ct) |
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Definition
# of PCR cycles to reach the point where fluorescence is detected.
a lower number means a higher concentration and vice versa |
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Term
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Definition
| FRET based, real-time PCR, hydrolyses probe, quencher interferes with fluor of report |
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Term
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Definition
1)Target DNA denatured to single strand
2) primer binds to template/target DNA
3)TaqMan probe binds
4) Taq polymerase synths DNA until reaches probe, it removes quencher so reporter can fluoresce |
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Term
| Molecular beacon method principle |
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Definition
| FRET based, realt- time PCR, hybridization probe, forms hairpin loop Q and R at either end |
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Term
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Definition
1) denatured, ends also denature, can now attach to target in annealing
2) Q and R separated by distance, R can now fluoresce
3) fluor increases as PCR product accumulated |
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Term
| SYBR green method principle |
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Definition
| dsDNA tags target still in the strand-real time PCR |
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Term
| Which tests are signal amplification? |
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Definition
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Term
| which tests are probe amplification? |
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Definition
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Term
| what is included in the cleavase invader? |
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Definition
| target DNA, probe, invader oligonucleotide, FRET cassette (includes fluor reporter, quencher, and cleavase enzyme) |
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Term
| What probes are used with bDNA? |
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Definition
| extender, preamplifier, and amplifier |
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Term
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Definition
1) cells lysed, RNAses inactivated
2) capture probes attached to tubes
3) sample RNA binds to these probes
4) extender, preamp, and amp
5) substrate added, light produces
6) measure amt of light, proportional to amt of target in sample |
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Term
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Definition
1) denatured to single strand
2) single strands hybridized to specific RNA probe
3) DNA-RNA hybrid captured by antihybrids
4) another anti hybrid added to tube
5) add substrate,light
6) measure amt of light |
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Term
| Which tests are target amplification? |
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Definition
| end point PCR, real time PCR, reverse transcriptase, multiplex PCR |
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Term
| Percent of DNA different between individual humans |
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Definition
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Term
| Random Match Probability calculation |
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Definition
locus frequency- 2pq
total RMP-frequencies multiplied together |
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Term
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Definition
| 8-90 bp long, noncoding, first discovered, inherited from both parents, good sample needed for DNA recovery, used in RFLP (southern blot) |
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Term
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Definition
| 1-8 bp, noncoding, inherited from both parents, degraded samples ok for DNA recovery, used in PCR, easier |
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Term
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Definition
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Term
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Definition
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Term
| single locus vs multi locus |
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Definition
| single-occurs once in genome, multi occurs multiple places |
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Term
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Definition
| 13 loci and amelogenin (indicated gender) |
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Term
| crime scene specimen collection |
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Definition
1) wear gloves (cells slough from skin)
2) tie hair back or cover it (one hair can implicate someone)
3) coughing or sneezing gets cells everywhere
4) don't accidentally transfer biologics |
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Term
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Definition
| circular, more stable than chromosomal, has hypervariable region, multiple copies, maternal inheritance, used in identity testing |
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Term
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Definition
| DNA treated with restriction enzymes, specific sequences ampd |
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Term
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Definition
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Term
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Definition
| 10 million markers, expensive |
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Term
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Definition
| only in males, inher from dad |
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