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What are the 4 DNA Isolation techniques?
What is DNA isolation?
A technique used to extract and separate DNA only from other cell components and foreign materials
What is Chelex Extraction used for and how does it work?
- Very small samples
- Uses chemical beads, chelex, that collect and bind all non-DNA materials
- Not good for RFLP
What is QiaAmp Extraction?
- Silica column bases method
- Cell/nucleus is lysed and rinsed
What is organic extraction?
Need Large Sample, but maintain quality and quantity better
What is differential extraction?
A technique to isolate DNA from other cell samples mixed with sperm cells
What is reverse dot blot PCR?
Probes are immobilized on a strip and PCR samples are added to the strip
What are the 3 steps of PCR?
What is the marker specific to male DNA?
What things can degrade DNA?
- UV light
What is star activity?
Non stringent condition for performance of specific enzyme
What is stochastic fluctuation?
Unequal representative of alleles due to sampling errors
What is Preferential amplification?
- One allele is amplified more than the other (looks homozygous)
- Allelic dropout
What is a heteroplasmy?
Mixture of samples in a single source
What is the purpose of DNA typing?
To identify if a person is the source of biological evidence
What are two typing systems for profiling?
- Detecting continuous alleles (RFLP)
- Detecting discrete alleles (PCR)
What are the 3 ways samples show genetic concordance?
- Samples are from the same source
- Coincidence (2 people have the same genetic profile for a particular marker)
- Error in collection and handling of sample
What two genetic models do we use to estimate allele frequency?
- Hardy-Weinberg Equilibrium
- Linkage Equilibrium
What does the frequency estimate?
- The evidence profile, not the suspect profile
- The evidence profile is then matched
What are the steps used in estimating profile frequency?
- Population studies (collect data)
- Frequency is tested for departure from H-W and LE
- Calculate Genotype frequency
- Calculate frequency by multiplication of loci
Why is it necessary to establish the DNA databank?
What is the difference between a databank and database?
- bank- all information about the offender and process of handling the case
- base- part of the databank
What is required for successful Databank?
- Sample collection
- Sample analysis
- Establishing a database for analyzed data