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Biochemistry BCC > L28: Recombinant DNA technology II > Flashcards

L28: Recombinant DNA technology II Flashcards

1
Q

What is polymerase

A

an enzyme which brings about the formation of a particular polymer, especially DNA or RNA.

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2
Q

Define Recombinant

A
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3
Q

An enzyme which catalyses the formation of a polymer

A

Polymerase

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4
Q

Allows selective sysnthesis of a gene sequence 100bp -10000bp in length

A

Polymerase chain reaction

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5
Q

Define: Polymerase chain reaction

A
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6
Q

The word _____ is derived from the Greek root poly-, meaning many, and mer,

A

Polymer

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7
Q

Say 5 things you know about Sanger Sequencing

A

1
2
3
4
5

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8
Q

define Sanger sequencing

A
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9
Q
A

Sanger Sequencing.

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10
Q

What is in vitro DNA synthesis in a tube?

A
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11
Q

What happens when you heat DNA pol to 95 degrees?

A

They will nature

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12
Q

What is ssDNA primer?

A
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13
Q

What in dNTPs

A

Deoxy…….

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14
Q

What is DNA polymerase?

A
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15
Q

What is ssDNA template?

A
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16
Q

How is ssDNA obtain?

A

Heat up to 95 degrees

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17
Q

What is Taq Polymerase?

A
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18
Q

What is thermus aquaticus?

A
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19
Q

What is an advantage to thermus aquaticus?

A

Withstands repeated heating and cooling cycles without denaturing

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20
Q

Why is mg2+ an essential cofactor in thermus aquaticus?

A
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21
Q

What reaction requires knowledge of gene sequence required to design ssDNA primers?

A

Polymerase chain reaction

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22
Q

Name 3 uses of Polymerase chain reaction uses

A
  1. Gene cloning: amplification of defined gene sequence
  2. VIral screening - high sensativity, detect virus before showing disease (covid)
  3. Forensics - DNA finger printing, show familial linkage, e,g OJ Simpson (DNA testing)
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23
Q

In PCR, what is primer pair selection?

A
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24
Q

How many primers are needed for dsDNA sysnthesis?

A

2

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25
Q

_____ ______ defines region to be amplified

A

Primer pair

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26
Q

What is a restriction enzyme?

A
27
Q

Restriction enzymes are not needed in any step of which kind of reaction?

A

Polymerase chain reaction.

28
Q

What is a DNA oligonucleotide?

A
29
Q
A

DNA oligonucleotide

30
Q

What does annealing mean?

A
31
Q

What is a primer annealing temperature?

A
32
Q

How many hydrogen bonds in A+T ?

A
33
Q

How many hydrogen bonds in C+G?

A
34
Q
A

ssDNA primer

35
Q

What is the process of PCR?

A

1.95 degrees -
2. 42-65 degrees -
3. 72 degrees -

36
Q

Define Taq polymerase

A
37
Q

define DNA synthesis

A
38
Q

What is DNA synthesis

A
39
Q
A

DNA synthesis

40
Q

In PCR, what is linear amplification?

A
41
Q

What are PCR cycles?

A
42
Q

What are PCR long v short DNA products?

A
43
Q

What is gel electrophoresis?

A
44
Q

What is Sanger DNA sequencing?

A
45
Q

What are dideoxyribonucleotides?

A

chain-elongating inhibitors of DNA polymerase, used in the Sanger method for DNA sequencing

46
Q
A

dideoxyribonucleotides?

47
Q

This was used to obtain the human DNA sequence

A

Sanger Sequencing

48
Q

What are the applications of Sanger Sequencing?

A
  1. Determine the order of baes from single gene to whole genome.
    2.Screens for gene mutations and sequence variants
  2. Validate the sequence of a PCR product
49
Q

Define: dideoxyribonucleotides chain terminators

A
50
Q

what are dideoxyribonucleotides? triphosphates? dNTPs

A
51
Q

Etymology of dideoxyribonucleotides?

A
52
Q

what is ddCTP?

A
53
Q

what is dideoxyATP

A
54
Q

what is dideoxyGTP

A
55
Q

what is dideoxyTTP

A
56
Q

what dieoxy ctp?

A
57
Q

what is ddATP

A
58
Q

what is ddGTP

A
59
Q

what is ddctp(?)

A
60
Q

What is ddNTP?

A
61
Q

What is ‘gel’ in DNA?

A
62
Q

What is gel electrophoresis?

A
63
Q

what are fluorescent ddNTP’s?

A
64
Q

what is a flourophore in sanger sequencing?

A

Biochemistry BCC (7 decks)

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