Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Bioc2201 > DNA cloning > Flashcards

DNA cloning Flashcards

1
Q

What is an insert?

A

Target DNA region that is incorporated into the vector.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

What are the two requirements for a vector?

A
  1. DNA molecule that can integrate foreign DNA.
  2. Can replicate it independently.
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

What are the three genetic features of typical cloning vectors?

A
  1. Origin of replication
  2. Multiple cloning site / polylinker.
  3. Selectable markers.
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

The MCS allows for independent replication. True or false?

A

False.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

The origin of replication facilitates the addition of the insert. True or false?

A

False.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

Antibiotic resistance genes are a common selectable marker. True or false?

A

True.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

In the vector, there is a short stretch of DNA with many unique RE sites. What is it called?

A

MCS / polylinker.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

What is the really common plasmid vector called?

A

pUC19.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What are the two selectable markers in the pUC19 vector?

A
  1. Ampicillin resistance gene.
  2. lacZ gene.
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Does transformation into host cells happen before or after ligation of the insert?

A

After.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

What enzyme carries out vector dephosphorylation?

A

Alkaline phosphatase.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

What type of enzyme carries out insert phosphorylation?

A

A kinase.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

What two things does vector dephosphorylation prevent?

A
  1. Self-ligation.
  2. Concatamerisation (different vectors bind).
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Insert phosphorylation is required for the activity of which enzyme?

A

T4 DNA ligase.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

What do we call cells that are more permeable to foreign DNA?

A

Competent.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

What chemical is used in chemical transformation?

A

Calcium chloride.

17
Q

What transformation technique uses short electric pulses?

A

Electroporation.

18
Q

If the insert is properly ligated, will lacZa be expressed?

A

No.

19
Q

We want a white phenotype in our cells. True or false?

A

True.`

20
Q

If beta-galactosidase is functional, do we have an insert-carrying vector?

A

No.

21
Q

What is the role of IPTG?

A

Induces the lac operon.

22
Q

If X-gal is not hydrolysed, what colour will the colony be?

A

White.

23
Q

Insertional inactivation will occur whether or not the insert is correct. True or false?

A

True.

24
Q

What is the main difference between a genomic and a cDNA library?

A

Genomic libraries contain all DNA, cDNA libraries contain only the expressed DNA.

25
Q

Lambda phages only infect E. coli. True or false?

A

True.

26
Q

What are the two structural components of a lambda phage?

A

A head (capsid) and a tail (sheath).

27
Q

What is a lytic cycle in viruses?

A

A replicative cycle where new phages are released via host lysis.

28
Q

What are three desirable features of lambda phages as vectors?

A
  1. dsDNA (REs can work)
  2. High infection rate.
  3. Large regions of non-essential DNA.
29
Q

What range of insert sizes can a lambda phage take?

A

15-21 kb.

30
Q

What total amount of DNA must the lambda phage have to exist?

A

46-52 kb.

31
Q

What is the lysate?

A

The culture of lysed bacteria and recombinant lambda phages.

32
Q

What do circular plaques on the bacterial lawn tell us?

A

E. coli cells have died.

33
Q

Each plaque represents a unique phage. True or false?

A

True.

34
Q

What is used to ‘copy’ the bacterial lawn?

A

A membrane/filter.

35
Q

Is the membrane treated with acidic or alkaline solution? Why?

A

Alkaline solution, to release DNA and denature it.

36
Q

DNA probes bind with dsDNA to identify the target insert. True or false?

A

False, ssDNA.

37
Q

Two possible ways to produce sticky ends for cDNA libraries?

A
  1. Restriction enzymes.
  2. Ligation of restriction site linkers.
38
Q

What enzyme is required for the first step of cDNA library creation?

A

Reverse-transcriptase.

39
Q

Bacteriophages are lysed on the membrane during lambda phage DNA cloning. True or false?

A

True.

Bioc2201 (11 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions