Biotechnology

Question 0

what is a vector?

Answer 0

molecules of DNA that can accept fragment of foreign DNA

Question 1

What is a restriction site?

Answer 1

A DNA sequence that can be cleaved by a restriction endonuclease

Question 2

what are the 3 characteristics of a vector?

Answer 2

must be capable of autonomous replication in cell
must carry at least one restriction site to insert foreign DNA
must carry at least one gene for selection (usually Ab resis)

Question 3

what is a genomic DNA library?

Answer 3

the entire genome is chopped up w/ restriction enzymes and cloned into vectors

Question 4

what is a cDNA library?

Answer 4

contains all the mRNAs present in a specific cell or tissue type at the time the mRNA was collected so you can see what genes are being expressed in that particular cell or tissue type

Question 5

if you wanted to take human DNA from a cDNA library and clone it using a bacterial vector, what would you need?

Answer 5

promoter

shine-Dalgarno sequence

Question 6

what ingredients do you need for DNA sequencing?

Answer 6

single stranded template
dNTPs
primer
polymerase

Question 7

what is special about dideoxynuleotides?

Answer 7

they are lacking a 3’ OH so when they are incorporated into a growing DNA strand, the synthesis terminates

Question 8

what is a probe?

Answer 8

ssDNA molecule that is labeled using radioactivity that can be hybridized to ssDNA that is complementary

Question 9

whta is the difference between small probes and large probes?

Answer 9

small probes= very specific

large probes= much less specific (don’t bind exactly)

Question 10

what is a genetic variation in a non-coding region with no disease?

Answer 10

polymorphism

Question 11

what is a genetic variation that causes disease in a non-coding region?

Answer 11

mutation

Question 12

An RFLP is present in a polymorphic region if…?

Answer 12

the genetic change creates or deletes a restriction site

the genetic change has more or less of a repeated sequence

Question 13

what are the 2 types of RFLPs?

Answer 13

SNP (single nucleotide polymorphism)

VNTR (variable number of tandem repeats)

Question 14

what are the 3 main steps to PCR?

Answer 14

denature
anneal primers
extend w/ DNA polymerase (Taq polymerase)

Question 15

what are the temperatures of each of the 3 steps of PCR?

Answer 15

annealing= 55C
Denaturization= ~95C
Extension= ~72C

Question 16

what are microarrays?

Answer 16

thousands of immobilized sequences (probes) on glass slides that are analyzed w/ a computer

Question 17

what is ELISA?

Answer 17

enzyme-linked immunosorbent assay
antigen is bound to the well of a microtiter plate and probed w/ antibody linked to an enzyme; color= positive rxn
looks for proteins

Question 18

what mutation and gene is involved w/ sickle cell anemia?

Answer 18

missense mutation

beta globulin gene

Question 19

what is unique about the disease causing mutation in sickle cell?

Answer 19

creates an RFLP not the other way around!

Question 20

what gene and mutation is involved w/ Cystic fibrosis?

Answer 20

CFTR gene

3 nucleotide deletion (missing phenylalanine)

Question 21

what is different about the mutation that causes PKU?

Answer 21

there are over 400 known mutations that cause it (in any of 13 exons)

Question 22

what is the gene and mutation associated w/ myotonic dystrophy?

Answer 22

trinucleotide repeat expansion in the 3’ NCR

protein kinase gene

Question 23

what is clinically important about myotonic dystrophy?

Answer 23

most common adult form of muscular dystrophy

Question 24

what is unique about testing for HIV?

Answer 24

we test for the antibodies, not the virus, so an incubation time is required in order to mount an immune response (~ 6 months)

Question 25

what can be used in both paternity testing and forensics as molecular fingerprinting?

Answer 25

VNTRs