chapter 10 - DNA Flashcards
James Watson and Francis Crick discovered
Developed DNA model and double helix structure in 1953
Rosalind Franklin discovery
Took first pictures of DNA
what is DNA
Code of genetic information
where is DNA found
nucleus
DNA helps code for
proteins
proteins give us
uniqueness
gene definition
fundamental unit of heredity
how many chromosomes does a person have?
46 total
23 pairs, 1 from each parent
chromosomes are found in the form of
wound up DNA
double helix
2 coiled strands of DNA
what are DNA strands made of?
nucleotides
nucleotide made of
5-carbon sugar (deoxyribose), phosphate group, nitrogenous base
4 base pairs of DNA and their pairing
adenine and thymine
cytosine and guanine
coding DNA
Codes for unique proteins
what percentage of DNA is coding DNA?
5%
what percentage of DNA is non-coding DNA?
95%
what does non-coding DNA code for?
nothing
what type of DNA is best for forensic identification? why?
non-coding DNA, contains tandem repeats that would be specific to each person
what is DNA replication
Making new DNA from existing DNA
Making more copies
how does DNA replication work?
- DNA helicase separates DNA strands by breaking Hydrogen bonds
- replication fork forms gaps in DNA ligase
- DNA polymerase adds complementary nucleotides to each DNA strand
- covalent bonds form along sugar-phosphate backbone
- DNA polymerase falls off
- replication is complete
what does the replication fork look like
Y
purpose of DNA fingerprinting
Establishes a link between biological evidence and a suspect
uses for DNA fingerprinting
Evidence in court
Body identification
Blood relative identification
Cures for disease
3 main technologies of DNA typing
RFLP
PCR
STR
what does RFLP stand for
Restriction Fragment Length Polymorphisms
what does PCR stand for
polymerase chain reaction
what does STR stand for
short tandem repeats
when was RFLP used
1985-mid 90s
polymorphisms definition
variations of DNA sequences between individuals
4 basic steps of RFLP
Isolate→ separating the DNA from the cell
Cut→use restriction enzymes to cut
Separate/Sort→ use gel electrophoresis
Analyze→ looking for specific allele patterns
what happens in the 1st step of RFLP?
isolation and extraction
Break open cells→grind them and add a detergent solution
Add salt→ (sodium chloride) - remove proteins bound to the DNA
Precipitate DNA→ add cold alcohol
what happens in the 2nd step of RFLP?
Cut
Restriction enzymes (bacterial cells) cut DNA
Looks for specific DNA sequence (tandem repeats)
what happens in step 3 of RFLP
Gel Electrophoresis
Load DNA into gel wells
Add electrical current
Molecules sort by size
Banding pattern
what happens in the 4th step of RFLP?
Radioactive markers are added
Bind to DNA
3 outcomes of RFLP
match
excluded
inconclusive
DNA match definition
DNA profile appears the same
DNA exclusion definition
samples originated from different sources
DNA inconclusive definition
doesn’t support a conclusion
what is PCR designed to do
Designed to copy or multiply strands of DNA
when is PCR used
when there is limited evidence
how much DNA can you get from a single speck of blood?
MILLIONS
primer definition
short segments of DNA
how does PCR work (4 steps)
Heat DNA→ causes separation of DNA strands
Cool and add a primer→ allows DNA segments to find their base pair
Add DNA polymerase and free nucleotides→ causes DNA to rebuild
Heat to 75 C for completion
advantages of PCR
Yields more than 1 billion copies
quick
short strands
anything can have DNA on it
how many cycles of PCR usually happens
28-32
how long does 1 cycle of PCR take
2 min
why are shorter strands of DNA more advantageous in PCR?
they are less susceptible to degradation
what is STR
Short sequences that repeat themselves in a DNA molecule
how long is an STR
3-7 bases in length; less than 450 bases long
where are STR located
chromosomes (loci)
what type of DNA testing is the most successful? why?
STR, large numbers in the human genome
how does STR work?
Extract the gene from the sample→ STR TH01→ A-A-T-G; 7 variants found→ 5-11 repeats
Amplify the sample using PCR
Separate by electrophoresis
Examine the distance the STR migrates to determine the number of times TH01 repeats
Each person has two STR types for TH01—one inherited from each parent
how are the results of STR shown?
peaks on a graph
each represents the size of a DNA fragment
the possible alleles are numbered for each loci
multiplexing definition
the more STR’s characterized, the smaller the percentage of population they can come from
2 forms of sex identification in STR
Amelogenin gene
Y-STR’s
Amelogenin gene
found on x and y chromosome
Shorter by 6 bases on the X chromosome
Males= 2 bands; females= 1 band
Y-STR’s
only found on Y chromosome
advantages of STR
only requires a small amount of DNA
greater discrimination of results
less susceptible to degradation
less time to run
how much DNA do you need for STR
18 cells