Flashcards in DNA recombinant II Deck (31):
if we are interested in amplifying a piece of RNA what do we use
reverse transcriptase to convert RNA into cDNA
When PCR continues normally we incorporate a dye what color is it and when does it start to glow
SYBR green dye, and it glows as more and more DNA is synthesized
DNA sequencing takes advantage of ddNTP'S what are they?
When we need to stop a DNA copy what do we use
how does a ddNTP work?
it inserts a deoxyribonucleotide with just a hydrogen bond and we need OH to polymerase
What do you need for DNA sequencing
2. template (what you want to sequence)
3. Normal dNTP'S
4. Enzyme and Buffer
and now a little ddNTP'S to stop DNA extension
What is the future what is Pyrosequencing?
that is when the two phosphate groups detach from the triphosphate group and react chemically with enzymes giving off a light energy that shows up on a pyrosequencing machine signifying that a bond has been made and dna has been replicated
In pyrosequencing what does each peak mean double height, single height and none
double height is when you have two or more of the same nucleotide. ex. ccc
single height is when only one nucleotide is added.
when there are none is because it did not match up with the template strand
what is the quality or state of being able to assume different forms?
polymorphism such as frogs
when one of two or more alternative forms of an allele from a chromosomal locus resulting from differences in DNA sequences or numbers of tandem repeats-not necessarily a gene. it can be anywhere
This is a known DNA sequence, often of DNA polymorphisms-best ones are unique in the genome, STS
DNA marker, sequence-tagged sites
What are the classes of DNA polymorphisms
SNP'S (single-nucleotide polymorphisms)
STR'S (short tandem repeats)
VNTR'S (variable number of tandem repeats)
What is southern blot?
analysis determines arrangement and location of restriction sites.
There are 3 blots what are their names and what are they used for?
how does southern blotting work?
dna is digested by enzymes producing fragments.
fragments are separated by size with gel electrophoresis.
dna fragments transferrred to a memebrane filter they are arranged exeactly as they are on the gel.
a labeled probe (radioactive or chemiluminescence) binds to DNA of interest.
This guy was a big player in the human genome project
J. Craig Venter
how many SNP'S were in J. Craig Venters diploid genome?
generally each gene is a STS- unique site in the genome
sequence tagged site
what is the most common DNA polymorphism caused by
a single point mutation of SNP'S
SNP'S can be harmful if they occur where? there are two places
coding genes, and regulatory regions
what does RFLP stand for?
Restriction Fragment Length Polymorphisms
what two things can be used to detect RLP'S
Southern blot or PCR
when were VNTR'S used to exonerate someone in a murder trial
what was the name of the murder trials that exonerated someone using VNTR'S
what are the positives for genetic testing
1. identifies mutations in key genes responsible for disease.
2. prenatal diagnosis
3. newborn screening-testing for genetic disease
4. carrier detection: testing of adults to calculate probability of baby having diseases-uses blood
why genetic testing doesn't always work
1. the gene for the disease hasnt been found.
2. the gene has many mutations making single test unreliable.
3. the presence of the gene does not always result in disease.
4. many diseases are polygenic, or caused by multiple genes
What takes advantage of DNA polymorphisms to identify individuals for forensics ?
most paternity tests use what to determine who the parents are
Gene therapy is mostly for what kind of cells
the procedure : take out cells, introduce normal gene (transgene) with virus vectors into them (now transgenic cell), replace in body is known as