BIotechnology (younger)

Question 1

Enzymes that exist in nature and cleave DNA into smaller, more manageable fragments.

Answer 1

Restriction endonucleases

Question 2

Restriction endonucleases recognize in general how many base pairs?

Answer 2

4 to 8 bp

Question 3

What is a short sequence of nucleotides that is read from 5’ to 3’ are the same on both strands?

Answer 3

Palindromes

Question 4

Which is a palindrome?
5’ TCTTCT 3’
3’ AGAAGA 5’

5’ TCTAGA 3’
3’ AGATCT 5’

Answer 4

bottom one is a palindrome

Question 5

What are sticky ends?

Answer 5

Restriction enzyme like a jagged/ staggered staircase cut and there is base pair overhang.

Question 6

What is a blunt end?

Answer 6

Restriction enzyme cuts straight through the DNA… There is no base pair overhang.

Question 7

True/False: Blunt and sticky ends both the 3’ OH group and the 5’ phosphate are attached after cleavage (important for ligation).

Answer 7

True

Question 8

DNA sequence that can be cleaved by a restriction enzyme is called a ___________.

Answer 8

restriction site

Question 9

True/False: the shorter the recognition sequence that restriction enzymes recognize the less bp you will have… and same with longer base pair recognition sites.

Answer 9

True, hence the longer the recognition sequence is, it determines how often it will cut.

Question 10

Fragments of DNA can be pasted together to make hybrid molecules called:

Answer 10

recombinant DNA

Question 11

What enzyme creates the phosphodiester bonds for DNA cloning?

Answer 11

DNA ligase

Question 12

True/False: DNA cloning involves inserting a restriction fragment into a ___________. A vector can be replicated in host cells (bacteria or yeast) and now DNA is cloned and amplified, this is called __________.

Answer 12

Cloning vector

Recombinant DNA amplification

Question 13

What are the most common vectors?

What are some others.

Answer 13

Prokaryotic plasmids

Bacteria and yeast

Question 14

What are the 2 different DNA libraries?

Answer 14

Genomic DNA libraries

cDNA libraries

Question 15

How are Genomic libraries made?
What do they represent?
How can they be used?

Answer 15

Genomic:
Could make a complete series of cloned end vectors and if you collect enough of the plasmids.
you could have an entire genome sequence and categorize them by where they are on the chromosome.

Question 16

How are cDNA libraries made?
What do they represent?
How can they be used?

Answer 16

(Complementary DNA)

If you collect the mRNA then you can reverse transcribe that and make DNA

Question 17

cDNA represents __________ DNA

Answer 17

expressed

Question 18

cDNA libraries do what?

Answer 18

allow one to see what genes were being expressed in a particular cell tissue type.

Question 19

cDNA only contains ‘_____’ sequences (no introns, promoters, etc.)

Answer 19

mRNA

Question 20

HBV doesn’t have organelles so thats why you use yeast cells… (IDK exactly what he was saying here) falling asleep…

Answer 20

look on slide 21 of biotech

Question 21

HBV doesn’t have organelles so thats why you use yeast cells… (IDK exactly what he was saying here) falling asleep…

Answer 21

look on slide 21 of biotech

Question 22

What do ddntps do? (important)

Answer 22

ddntp stop elongation… they don’t have OH group so chain elongation ends and it determines where it stops on the gel or where the growing chain stops. Those nucleotides are also used in cancer treatments.

Question 23

What is a piece of ssDNA and is used to identify DNA fragments (usually labeled using radioactivity)?

Answer 23

Probes

Question 24

You have 3 people…You know that one person has sickle cell, one doesn’t have it, and you have another person you want to see if has sickle cell mutation… what do you do?

Answer 24

Take different samples generated from gpcr of the 3 peoples DNA, take the probe and bathe it, if probe binds to person with sickle cell and not with another person who doesn’t have it. and the person that you are wondering about, the probe binds then you know they have sickle cell.

Question 25

What are the different types of probes?

Answer 25

smaller probes- chemically synthesized oligonucleotides (diagnostic)
Larger probes are comparative- looking for comparisons among different organisms.

Question 26

What is an analysis of DNA?

Answer 26

Southern Blotting

Question 27

What is an analysis of mRNA?

• only detects expressed sequences
• used for tissue or cell specific studies
• quantitative

Answer 27

Northern Blotting

Question 28

What is an analysis of Protein?

• antibody specific to the protein of interest
• also quantitative and qualitative in nature

Answer 28

Western Blot

Question 29

Humans are not related, why?

Answer 29

Regions not part of expression are modified and changed in hypervariable regions.

Question 30

Genetic changes that result in a disease is called what?

Answer 30

Mutation

Question 31

Genetic variations in non-coding regions with no disease are called ________.

Answer 31

Polymorphisms

Question 32

If you have a disease causing genetic variation that is handed down from generation to generation it is what?

Answer 32

slide 31 have to listen to get answer

Question 33

RFLP means what?

Answer 33

Restriction Fragment Length Polymorphism

Have used restriction enzymes to cut DNA, length of DNA fragments are of different length due to polymorphism..

Question 34

If a genetic change in a polymorphic region one of 2 things had to have happened… What are those 2 things that makes a RFLP be present?

Answer 34

Creates or deletes a restriction site

has more or less of a repeated sequence

Question 35

What do VNTR’s do?

Answer 35

Paternity test to determine heredity… Also CSI
Varies greatly from non related individuals
Area that contains many tandem repeats

Question 36

What may create or abolish a restriction site?

Answer 36

SNPs (Single nucleotide polymorphisms)

Question 37

RFLPs do what to a restriction site?

Answer 37

RFLP is either caused by SNP that results in creating or abolishment of a restriction site
or caused by VNTRs that are just tandem repeats