Brainscape's Knowledge GenomeTM


Top Flashcards (Certified)
All Flashcards

Molecular Biology Lecture > 3: DNA Sequencing Techniques (MIDTERMS) > Flashcards

3: DNA Sequencing Techniques (MIDTERMS) Flashcards

1
Q

These two methods in sequencing DNA became the basis of more sophisticated DNA techniques

A

Sanger and Maxam-Gilbert Technique

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

What are the three DNA sequencing discussed?

A
  1. Sanger Technique
  2. Maxam-Gilbert Technique
  3. Next Generation Technique
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

Technique made during the mid 1970’s

a. Sanger Technique
b. Maxam-Gilbert Technqiue
c. Both
d. NOTA

A

c. Both

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

Process of determining sequence of nucleotide bases

A

DNA Sequencing

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

How are nucleotides identified?

A

Based on the bases of their structures

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

Use this card to familiarize the process of DNA sequencing

A
  1. Break down DNA to smaller pieces
  2. The smaller pieces will be sequenced one at a time
  3. That portion of DNA will assemble the sequence into a single continuous order of nucleotide
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

T or F

Portion of DNA will assemble the sequence into a single continuous order of nucleotide to produce a sequence of genome

A

T

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

Conventional methods

a. Sanger Technique
b. Maxam-Gilbert Technqiue
c. Both
d. NOTA

A

c. Both

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

Sanger and Maxam-Gilbert Technique are the principle of what new sequencing method?

a. Robotics
b. Automated Sequencing
c. Both
d. NOTA

A

c. Both

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Chain termination sequencing

a. Sanger Technique
b. Maxam-Gilbert Technqiue
c. Both
d. NOTA

A

a. Sanger Technique

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

What DNA sequencing technique:

→ Target DNA is copied many times
→ PCR-based techniques

A

Sanger Technique

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

T or F

Sanger Technique makes fragments of different lengths

A

T

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

→ 1991-2003

→ Process wherein the entire human genome or makeup of a human being is sequenced

A

Human Genome Project

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

When was the human genome project started and ended?

A

1991-2003

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

Technique wherein results are produced within days

a. Sanger Technique
b. Maxam-Gilbert Technique
c. Both
d. NOTA

A

d. NOTA (next generation sequencing technique yarn)

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

Uses fluorescence

a. Sanger Technique
b. Maxam-Gilbert Technique
c. Next generation technique
d. ALL of the above
e. NOTA

A

d. ALL of the above

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

What DNA sequencing technique:

→ PCR-based method

→ Modified DNA replication method

→ Growing chains are terminated by dideoxynucleotides/ddNTPs

A

Sanger Technique: chain termination method

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

What are the five requirements in Sanger Technique?

A
  1. DNA primer
  2. DNA polymerase
  3. dNTP
  4. four ddNTPs
  5. Template DNA
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

Sanger Technique:

What is the 1st step?

A

DNA Denaturation

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

Sanger Technique:

What is the 2nd step after denaturation?

A

Aliquot to 4 tubes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

Sanger Technique:

What is the 3rd step after aliquoting to 4 tubes

A

Incorporate reagents: Primer, dNTP, DNA Polymerase

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

Sanger Technique:

What are the reagents added before ddNTPs?

A

Template ssDNA, Primer, dNTP, DNA Polymerase

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

Sanger Technique:

After incorporating reagents, what should be added to the 4 tubes?

A

Four dideoxynucleotides or ddNTPs in each tube

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

Sanger Technique:

T or F

In adding ddNTPS in tube, 2 ddNTPS should be added per tube

A

F (1 specific ddNTP should be added per tube; example: Tube 1 - ddATP → Tube 2 - ddTTP)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
Sanger Technique: What is the last step after adding ddNTP?
Gel electrophoresis
26
How is the gel electrophoresis product of Sanger technique read?
BOTTOM to TOP and individually (tama nman khit sa kama rin eme)
27
Sanger Technique: In gel electrophoresis, DNA fragments move from?
negative to positive pole
28
Sanger Technique: T or F DNA fragments move from negative to positive pole because of the base backbone
F (PHOSPHATE backbone which is negative charged)
29
Sanger Technique: Identify what size of fragment → Migrate at the bottom → lighter → Read first
Shorter fragments
30
Sanger Technique: Identify what size of fragment → Migrate at the top → Heavier
Longer fragments
31
Sanger Technique: Migrate at the bottom and is read first a. Longer fragments b. Shorter fragments c. both d. NOTA
b. Shorter fragments
32
Sanger Technique: After gel electrophoresis, what is the product formed?
Complementary strand
33
→ Has OH on 3’ carbon → Allows attachment of nucleotides → Has more concentration
dNTP
34
→ Lacks OH on the 3’ carbon → Terminates synthesis of new strands → Fluoresce → Added in chain randomly → complementary to target DNA
ddNTP
35
Allows attachment of nucleotides a. dNTP b. deoxynucleotide triphosphate c. both d. NOTA
c. both
36
Has OH on 3’ carbon a. ddNTP b. dideoxynucleotide triphosphate c. both d. NOTA
d. NOTA (dNTP/deoxynucleotide triphosphate)
37
Terminates synthesis of new strands and fluoresce a. ddNTP b. dNTP c. both d. NOTA
a. ddNTP
38
Added in chain randomly and complementary to target DNA a. dNTP b. dideoxynucleotide triphosphate c. both d. NOTA
b. dideoxynucleotide triphosphate
39
T OR F ddNTP has more concentration than dNTP
F (reverse; dNTP pa rin lamang sa concenctration)
40
→ DNA sequencing method based on chemical modification of DNA and subsequent cleavage at specific bases → Chemicals are used that selectively attack purines and pyrimidines
Maxam-GIlbert Technique: Chemical Sequencing
41
Use this card to familiarize the process of Maxam-Gilbert Technique
1. Denature DNA 2. Chemical modification of DNA/Labelling 3. Purification of the DNA fragment to be sequenced 4. Chemical treatment generates breaks in DNA 5. Addition of Piperidine 6. Gel electrophoresis
42
Maxam-GIlbert Technique: What is the 1st step
Denature the DNA
43
What is the temperature for denaturing DNA?
95 degree celcius
44
Maxam-GIlbert Technique: After denaturing DNA, what is the next step?
Chemical modification aka radioactive labeling at one 5’ end of the DNA by kinase reaction using gamma -32 P ATP
45
Maxam-GIlbert Technique: In labelling the strand, what should be used?
gamma -32 P ATP
46
Maxam-GIlbert Technique: T or F Radioactive labeling at one 3’ end of the DNA by kinase reaction using gamma -32 P ATP
F (Radioactive labeling at one 5’ END of the DNA by kinase reaction using gamma -32 P ATP)
47
Maxam-GIlbert Technique: T or F Purpose of radioactive labelling is to mark the strand before cutting so that there are three 5'end
F (mark the strand before cutting so that there are TWO 5'end)
48
Maxam-GIlbert Technique: After radioactive labelling, what is the 3rd step?
Purification of the DNA fragment to be sequenced
49
Maxam-GIlbert Technique: After purification, what is the next step?
Chemical treatment generates breaks in DNA
50
Maxam-GIlbert Technique: What is the Chemical Process, Nucleic Acid Modifier, and Minutes Reaction Time for the base C?
Maxam-GIlbert Technique: Chemical Process: Splits C rings Nucleic Modifier: Hydrazine + Salt Reaction Time: 8 mins
51
Maxam-GIlbert Technique: What is the Chemical Process, Nucleic Acid Modifier, and Minutes Reaction Time for the base T + C?
Chemical Process: Splits pyrimidine rings Nucleic Modifier: Hydrazine Reaction Time: 8 mins
52
Maxam-GIlbert Technique: What is the Chemical Process, Nucleic Acid Modifier, and Minutes Reaction Time for the base G + A?
Chemical Process: Protonates purines Nucleic Modifier: Formic acid Reaction Time: 5 mins
53
Maxam-GIlbert Technique: What is the Chemical Process, Nucleic Acid Modifier, and Minutes Reaction Time for the base G?
Chemical Process: Methylates G Nucleic Modifier: Dimethylsulphate Reaction Time: 4 mins
54
Maxam-GIlbert Technique: Hydrazine + salt a. C b. T + C c. both e. NOTA
a. C
55
Maxam-GIlbert Technique: Hydrazine a. G + A b. G c. both e. NOTA
e. NOTA (T + C or PYRIMIDINES)
56
Maxam-GIlbert Technique: Formic Acid a. G + A b. G c. both e. NOTA
a. G + A
57
Maxam-GIlbert Technique: Dimethylsulphate a. G + A b. G c. both e. NOTA
b. G
58
Maxam-GIlbert Technique: Protonates a. G + A b. G c. both e. NOTA
a. G + A
59
Maxam-GIlbert Technique: Methylation a. G + A b. G c. both e. NOTA
b. G
60
Maxam-Gilbert Technique: Splits rings a. C b. T + C c. both e. NOTA
c. both
61
Maxam-GIlbert Technique: Splits C rings a. Hydrazine b. Hydrazine + salt c. both e. NOTA
b. C
62
Maxam-GIlbert Technique: Splits purine rings a. T + C b. C c. both e. NOTA
e. NOTA (there's no splitting of purine only PYRIMIDINE by using hydrazine)
63
Maxam-GIlbert Technique: Minutes reaction time: 8 minutes a. T + C b. C c. both e. NOTA
c. both
64
Maxam-GIlbert Technique: After treating with chemical, what is the next step?
Addition of Piperidine
65
Maxam-GIlbert Technique: This cuts sequence of the nucleic acid modifier
Piperidine
66
Maxam-GIlbert Technique: After addition of Piperidine, what is the last step?
Gel electrophoresis
67
Maxam-GIlbert Technique: Gel electrophoresis of Maxam-Gilbert Technique is read from?
BOTTOM to TOP, by PAIR
68
Read from bottom to top a. Sanger Technique b. Maxam-Gilbert Technique c. both d. NOTA
c. both
69
→ also called massively parallel sequencing → methods in which millions of DNA templates are sequenced simultaneously in a single reaction.
Next Generation Sequencing
70
Next Generation Sequencing: nucleotide sequence data is then assembled using ?
computer algorithms into a continuous genome sequence.
71
Next Generation Sequencing: What is the 1st step?
Ligation: fragments of cellular DNA are ligated to oligonucleotides referred to as adapters.
72
Next Generation Sequencing: What are the fragments of DNA ligated to oligonucleotides called?
adapters
73
Next Generation Sequencing: After ligation with adapters, what is next step?
Denaturation and strands are captured on solid surface
74
Next Generation Sequencing: Which serves as primer for DNA polymerase?
annealed oligonucleotide
75
Next Generation Sequencing: T or F After DNA extension, two long strands separate, leaving 1 strand attached by a covalent bond to a fixed oligonucleotide.
T
76
Next Generation Sequencing: Can form a bridge with another fixed oligonucleotide in another round of PCR proceeds.
DNA fragment
77
Next Generation Sequencing: After multiple rounds, what forms in the location of each original cellular fragment
a small patch of DNA
78
Next Generation Sequencing: T or F These single-stranded DNA molecules represent two sequences that are complementary to each other
T
79
Next Generation Sequencing: T or F Nucleotides contain a starting group on their 3’ ends
F (Nucleotides contain a BLOCKING group on their 3’ ends)
80
Next Generation Sequencing: induces the nucleotide to fluoresce and the color is recorded
Laser
81
Next Generation Sequencing: T or F The cycle can be repeated up to 20x only
F (repeated up to 100 X)
82
Next Generation Sequencing: assemble overlapping sequence fragments into longer pieces and in this way determine the overall sequence of a genome
Software packages
83
IF YOU SEE THIS CARD, PAKIREVIEW ANG STEPS NG NEXT GEN SEQUENCING PLS AND HIRAP NIYA GAWAN NG CARD FR
DALI NA PLS BB : (((
84
Denaturation Step a. Sanger b. Maxam-GIlbert c. Next Generation sequencing d. ALL e. NOTA
d. ALL
85
DNA Amplification and Extension a. Sanger b. Maxam-Gilbert d. ALL e. NOTA
a. Sanger
86
Ligation a. Sanger b. Maxam-GIlbert c. Next Generation sequencing d. ALL e. NOTA
c. Next Generation sequencing

Molecular Biology Lecture (17 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2025 Bold Learning Solutions. Terms and Conditions