Brainscape's Knowledge GenomeTM


Top Flashcards (Certified)
All Flashcards

MCBG (Milly's) > Session 12+14 (Molecular Techniques) > Flashcards

Session 12+14 (Molecular Techniques) Flashcards

(50 cards)

Start Studying
1
Q

What is needed for protein gel electrophoresis? (4)

A

Gel, buffer (maintains charge on sample), power, stain

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

What type of electrophoresis uses proteins intrinsic properties?

A

Serum protein electrophoresis

density of band = height of graph

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

What type of electrophoresis separates proteins according to size? How?

A

SDS-PAGE

Secondary and tertiary structures removed by beta-ME and SDS adds a negative charge to strand
Simply looking at polypeptides

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

What type of electrophoresis separates proteins according to charge? How?

A

IEF (isoelectrical focusing)

Gradient of pH is set up (low pH/+charge. High pH/-charge), protein moves until they reach a pH=pI then movement stops because they have no net charge

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

What type of protein gel electrophoresis separates proteins according to size and charge?

A

2D-PAGE

Charge (horizontal movement)
Size (vertical movement)

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

When would you use 2D PAGE?

A

When you have complex mixtures (ie whole cells)

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

What is the process of protein identification called?

A

Protenomics

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

What is an epitope?

A

A few amino acids on a protein that antibodies bind to

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

What are the two main types of antibodies?

A

Polyclonal - binds to many epitopes

Monoclonal- 1 epitope

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

When would you use antibodies as probes?

A

Western blotting

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

What does ELISA mean?

A

Enzyme linked immunoabsorbant assay

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

What are the 3 stages of PCR?

A

Denaturing
Annealing
Extending

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

What are the temperatures for the different stages of PCR?

A

Denaturing- 95
Annealing - 60
Plolymerising - 72

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

What builds the new strands on DNA in PCR?

A

DNA Taq polymerase

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

What sort of primers are needed for PCR?

A

Forward and reverse

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

What are the uses of PCR?

A

Amplify fragment
Investigate single base mutations
Investigate small deletion/insertions
Investigate Variation in genetic relationships

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

What enzymes cut DNA at specific palindromic sequences?

A

Restriction endonucleases

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

Why are DNA fragments negatively charged?

A

Phosphate group on nucleotides (PO4)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

What is the positive electrode called?

A

Anode

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

What do you need for DNA gel electrophoresis? (4)

A

Gel
Buffer
Power supply
Stain

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

What are the uses of DNA gel electrophoresis? (3)

Study These Flashcards
A

Investigate size of DNA fragments
Investigate mutations
Investigate DNA variation

22
Q

What is a plasmid + isolated gene to be copied called?

Study These Flashcards
A

Recombinant plasmid

23
Q

What is the vector for cloning DNA usually?

24
Q

What is used to reseal plasmids?

Study These Flashcards
A

Ligase enzymes

25
Why clone human genes? (4)
Make useful proteins Find out what genes do Genetic screening Gene therapy
26
What do you use to get DNA from mRNA?
Reverse transcriptase
27
What gene is isolated when making human insulin?
Proinsulin (immature form of the gene)
28
Why can gel electrophoresis be used to separate molecules of different size?
Large DNA fragments move slowly, smaller fragments move faster
29
What do restriction enzymes cut?
Phosphodiester bonds
30
What is DNA sequencing?
Process of determining the precise order of nucleotides within a DNA molecule
31
How is DNA sequencing done?
Heat DNA + primer + dideoxynucelotides (tagged) Chains grow Separate out new strands by gel electrophoresis Use computers and laser to read
32
Why are dideoxynucleotides used?
They lack an OH group meaning elongation is terminated
33
In PCR, at what end does the primer attach and why?
The 3' end because the DNA chain grows in a 5'-3' direction
34
What are allele specific primers?
Sequences that allow for a change in sequence and will bind accordingly
35
What is southern blotting?
Separating DNA by gel electrophoresis Transfer to nylon membrane (Done by soaking gel in alkaline solution- creates ssDNA) Mix with DNA probes (dont need 100% complementary) Detection methods detect binding
36
Why use southern blotting?
Allows for detection of difficult to find DNA Can be used in conjunction with PCR to detect: Gene structure expansion and repeats, mutations, genetic variation
37
How is northern blotting different to southern blotting?
It uses RNA instead of DNA
38
Why is northern blotting more difficult than southern blotting?
RNA degrades
39
What sort of primer would you use for RT-PCR? Why?
One with many TTT's @ 5' end so it can bind to the polyA tail at the 3' end of mature RNA
40
What do you compare during microarray?
2 'conditions'
41
How many DNA sequences do you look at at once in microarray?
Thousands
42
What does microarray allow us to see?
Gene expression in an individual
43
What is microarray?
``` Putting all genes on plate, Take someone's mRNA (from tissue) Use RT to produce cDNA Label cDNA Wash cDNA over plate cDNA will bind to genes Therefore gene expression can be seen ```
44
What are mini-satellites?
Repeating areas of base pairs (Can have the same repeat multiple times at a single loci) (Variable between homologous chromosome pairs)
45
How does DNA fingerprinting work?
Use restriction enzymes to chop up DNA Complete southern blotting Compare runs
46
What section of DNA are you looking at in DNA fingerprinting?
The non coding regions of the DNA
47
What are the probes used for DNA fingerprinting?
Minisatellites
48
What is karyotyping?
Lining up of chromosomes
49
What does FISH stand for?
Fluorescent in situ hybridisation
50
How do you do a FISH?
Make specific DNA sequence copy, Label probe Denature DNA and allow hybridisation Section of DNA becomes visible

MCBG (Milly's) (15 decks)

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