DNA CLONING Flashcards
1
Q
Characteristics of a cloning
vectors
A
- It must be self-replicating inside host cell, by presence of (Ori). -It must possess restriction site (MCS) for RE enzymes -Introduction of donor DNA fragment must not interfere with replication property of the vector - It must possess some marker gene such that it can be used for later identification of recombinant cell.
2
Q
For isolation of recombinant cell from nonrecombinant cell, ___________ is
employed.
A
marker gene of plasmid vector
3
Q
PBR322 plasmid vector contains different
marker gene?
A
Ampicillin resistant gene and Tetracycline Resistant
4
Q
ScaI is ____ reisistant
A
Ampicillin
5
Q
Lac gene works with X-gal producing ____ colonies
A
Blue
6
Q
LacZ + EcoR1 =
A
White colonies
7
Q
Steps of polymerase chain reaction?
A
- Denaturation (95 C)
- Annealing (60C)
- Extensions (72C)
8
Q
Required Temperature for denaturation?
A
94C
9
Q
PCR components?
A
- DNA Template
- Primer
- Taq polymerase
- Nucleotides
- Buffer (MgCl2)
10
Q
Why is Taq polymerase isolated from Thermus Aquaticus
A
Thermophillic Eubacterium found in hot springs
11
Q
Short synthetic oligosaccrides responsible
A
Primer
12
Q
PCR Application?
A
- Diagnosis Of Infectious Disease
- Forensic Medicine
