biotechnology
Why are "sticky ends" of DNA sticky?
"Sticky ends" contain a single-stranded end complementary to the target fragment's single-stranded end with which it can pair.
what is a western blot
. A technique in which proteins are separated by electrophoresis and probed with a specific antibody
what year did kary mullis discover the pcr
1983
what year did kary mullis win the nobel prize in chemistry
1993
15. What is the optimal temperature for the Taq polymerase to function??
720C
what do dideoxynucleotides do
Acts as a chain terminator when incorporated into the DNA because it lacks an OH at the 3' position.
what does the pcr do
Allows the amplification of a single DNA fragment from a small amount of a complex mixture of DNA
12. Which technique is used to identify suspects at crime scenes?
DNA fingerprinting
what is gel electrophoresis used for
DNA fingerprinting and sequencing
When "sticky ends" of two DNA molecules are paired, their sugar-phosphate backbones can be permanently rejoined by
DNA ligase.
9. What is needed for both DNA replication and PCR?
DNA template and DNA polymerase
8. List the three steps of PCR and what is occurring at each.
Denaturation- heat to uncoil DNA Annealing of primers- addition of primers to start replication Synthesis- addition of new nucleotides
the most common bacterium used in clonin is
E. coli
14. Name the bacteria the is used frequently in biotechnology techniques.
Escheria coli...E coli
3. Which process separates DNA using electricity?
Gel electrophoresis
what are the three types of restriction enzymes
I, II, III
4. Which enzyme seals strands of DNA back together?
Ligase enzyme
7. Which process is used to make many copies of DNA so it can be studied and experimented with?
PCR, Polymerase Chain Reaction
what are restriction fragment length polymorphisms
Point mutations found in a population that can eliminate or create restriction sites.
10. Name the enzyme that was isolated from a retrovirus and is used to make cDNA?
Reverse transcriptase
11. Which enzymes are needed to build cDNA library?
Reverse transcriptase and DNA ligase
where are restriction endonucleases found
bacterial cells
what other type of end does type ii make
blunt ends
16. How is cDNA different from a regular genomic library?
cDNA does not contain introns
Which statement best explains the role of a vector in recombinant DNA technology?
carries a segment of recombinant dna
what process occurs in the nucleus, uses helicase, involves okazaki fragments, and produces a lagging strand
cellular dna replication
what does cDNA stand for
complimentary dna
what are the three steps o fpcr
denature dna, annealing of primers, synthesis
what can dna fingerprints help do
diagnose genetic disease, biological relationship, identify missing person, identify crime scene suspects
what is the dna library
dna fragments from a source
what are the needed materials for the pcr to occur
dna to be copied, DNA polymerase...Taq polymerase, four dna molecules, two primers
what type of field is the dna place in in gel electrophoresis
electrical field
13. On what bases are DNA fragments separated during gel electrophoresis?
fragment size, number of base pairs
5. When are Okazaki fragments seen?
fragments produced during DNA replication, not part of molecular techniques
The technique used to separate DNA molecules based on their length/mass is called:
gel electrophoresis.
Which statement best explains why DNA fragments can be separated using gel electrophoresis?
have a negative charge
what temperature does the denaturing of dna take place in
high
what is the dna library introduced into
host cells
what does the western blot do
identify protein, antibodies
DNA fingerprinting would be a useful technique for:
identifying remains following natural disasters. distinguishing between potential suspects in a crime. reuniting families separated by the Holocaust.
waht does the northern blot do
indentify rna
what is a southern blot used for
indentifying a fragment of dna by radioactive label or flourescent dye
what happens to the dna library
inserted into plasmid vector, introduced into host cells
what temperatures does synthesis take place in
intermediate
who discovered teh polymerase chain reaction
kary mullis
what enzyme is used to attach dna strands
ligase
what temperature does the annealing of primers take place in
low
what makes cDNA different from other DNA libraries
made from mRNA
what happens in short tadum repeats
mutations change non coding sequences
what type of charge does dna have
negative
what type of DNA fragments can sticky ends easily rejoin with
others with sam ends
where does type ii of restriction enzymes cut
palindromes
what process occurs outside the nulceus, requires denaturization, requires heat cycles, and uses taq polymerase
pcr
what is the dna library inserted into
plasmid vector
what gel is the dna placed in in gelelectrophoresis
porous agarose gel
what pole does the dna migrate toward
positive
what are restriction endonucleases also known as
restriction enzymes
what is dna cut into fragments by
restriction enzymes
What are restriction endonuclease?
restriction enzymes that cleave (cut) DNA
what did restriction endonucleases create
restriction map, identification, DNA molecules to experiment with, help create recombinant molecules
what is a section of dna recognized by a restriction endonuclease
restriction site
what is an enyme used in cDNA
reverse transcriptase
what does gel electrophoresis do
separates dna pragments
2. What are sticky ends?
single-stranded complementary tails produced by restriction enzymes
which moves more quickly: large or small DNA fragments
small
6. Describe plasmids.
small, circular extra chromosomal DNA found in bacteria
what are sticky ends
staggered cuts
restriction endonucleases that produce what are prefered for recombinant dna technology
sticky ends
Why is DNA fingerprinting possible?
the number of repeats in a tandum sequence is highly variable from one individual to another
what did bacteria use restriction endonucleases for
to cute invading viruses
17. What is a Southern blot used for?
to identify a specific DNA sequence
18. What are RFLPs used for?
to identify an individual using a particular gene as a marker