Brainscape's Knowledge GenomeTM

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


See full index

MCBG > MCBG 13+14 - Molecular techniques > Flashcards

MCBG 13+14 - Molecular techniques Flashcards

1
Q

How does hybridisation work? 3

A
  1. ) Denature ds DNA
  2. ) Add a ssDNA or RNA probe (sequence of Nucleotides)
  3. ) Eithe probe is fluorescent/radioactive or you add a label
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Why was it named southern blotting?

A

Discovered by Prof Sir Southern

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

What is a Southern/Northern Blot. One word

A

Hybridisation. (Gel electrophoresis + hybrisation)

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

How is western blot different from southern blot?

A

Its uses proteins

DOES NOT USE HYBRIDISATION

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

Describe Western blot

A
  1. Gel Electrophoresis
  2. Blotting onto surface
  3. Adding labelling probs (antibodies) No hybrisation
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

Describe Southern Blot

A
  1. Restriction enzymes cleave DNA
  2. Gel Electrophoresis
  3. Transfer to nylon
  4. Hybridise with gene probe ( fluorescence or radioactive)
  5. Detect labels
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

Sickle cell disease is a mutation in the HBB
gene. What is the most suitable way to get the
new-born’s HBB gene for a genetic test?

A

PCR (part of ) new-born’s HBB

gene

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

After you PCR what can you then do after? 2/3

A
  1. restriction analysis
  2. gel electrophoresis (Southern blotting)
  3. DNA sequencing
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What is sickle-cell disease?

Whats the most common?

A

Group of blood disorders with the most common being sickle cell annemia.

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

MstII is a restriction enzyme; where are restriction

enzymes found?

A

Bacteria

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

Whats a single-nucleotide polymorphism?

A

A different base. A specific base position in the human genome, the base C may appear in most individuals, but in a minority of individuals it doesnt

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

Name three diseases single-nucleotide polymorphism occur in?

A

CF
Sickle Cell Disease
Beta Thalassemia

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

What units make up haemoglobin

A

HbA = a2b2

Alpha and beta subunits

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

Sickle Cell Anaemia. What mutation occurs?

A

Single Base mutation(exchange).
A->T in the B-globin gene
Glutamate -> Valine

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

Sickle Cell Anaemia. Why does the mutation effect RBCs?

A

Valine is hydrophobic. This distorts the shape of the cell. Causes it to collapse and form a sicle shape

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

Sickle Cell Anaemia. What is the name of the of Mutation (found on a biomed form) [hint aa]

A

Glu6Val

or E6V

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

What type of genetic disease is it?

A

Autosomal recessive

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

Whats a mismatch mutation?

A

C=T

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

Whats a pyrimidine dimer?

A

MUTATION

Where two pyrimidines (C,U,T) bond

20
Q

Whats an intercalating agent

A

hidium bromide
proflavine
molecules that may insert between bases in DNA, causing frameshift mutation during replication

21
Q

Name sources of mutations

4/5

A
  1. Ionising radiation
  2. UV light
  3. Oxidising chemicals
  4. inflammation
  5. Smoking
22
Q

How is DNA read in transcription?

A

DNA is read 3’ to 5’

23
Q

What direction is RNA synthesised in transcription?

A

mRNA synthesis 5’ to 3’

24
Q

What direction is mRNA read translation?

A

mRNA is read 5’ to 3’

25
Q

What direction is polypeptide chains synthesised?

A

polypeptide synthesis N to C

REVERSE ALPHABETICAL

26
Q

Single nucleotide changes can be split into two types.

A

Transition - Purine to purine

Transversion - (A/G-C/T)

27
Q

Whats a missense mutations?

A

A mutation that causes any amino acid to be changed from its original

28
Q

Whats a non-sense mutations?

A

A mutation that causes a premature stop codon

29
Q

Whats a silent mutation?

A

A mutation that doesnt effect the final mutation

30
Q

Whats a conservative mutation?

A

A mutation that changes an amino acid to one that has similar properties

31
Q

Whats a non-conservative mutation?

A

A mutation that changes an amino acid to one that has different properties

32
Q

What type of mutation is sickle cell anaemia? Sense missense or non-sense

A

Non-conservative MISSENSE Mutation

(a point mutation that doesnt cause a FS but changes the amino acid GLU-> VAL. So its a MISSENSE Mutation

33
Q

Missense mutations come in two different types

A

conservative mutation

non-conservative mutation

34
Q

IMPORTANT.

What are the three (that i care about) Classification of mutations?

A
  1. ) Effect on structure
  2. ) Effect on protein sequence
  3. ) Nomenclature
35
Q

In the classification that explains an effect on structure. What types of mutations are there? 6 (3+3)

A 
Micro:
Insertion
Deletion
Substitution
Macro:
Duplication
Deletion
Translocation
36
Q

In the classification that explains an effect on protein sequence. What types of mutations are there? 4

A

Frame Shift
Missense (Conserve and Non-Conserve)
Nonsense
Silent

37
Q

In the classification that explains nomenclature. What types of mutations are there? 3

A 
Nucleotide Substitution (76A>T)
Amino acid Substitution (E6V)
Amino acid deletion (/\F508)
38
Q

How does DNA sequencing work? 4

A
  1. )Take DNA and Amplify the sample sequence using PCR (then denature it)
  2. )Add Bases, DNA polymerase and fluorescently labelled di-deoxyribose nucleotides
  3. ) Gel electrophoresis
  4. ) Analyse using a computer and laser
39
Q

How does DNA finger printing work (in comparision to sequencing). Basically what are the steps? 3

A

It takes VNTRs (variable number tendem repeats) STR (short tandem repeats) amplifies them and then gel electrophoresis.

40
Q

What is reverse transcriptase PCR

A

Type of PCR

Uses mRNA instead of DNA and Reverse transcriptase enzymeto convert it into cDNA

41
Q

How do microarrays work?

A

Extract either DNA or mRNA from a cell (you tend to go for one normal and one diseased cell). If mRNA you need to reverse transcriptase PCR it and ad a lable. The mix of labelled GM is added to the microarray where HYBRIDISATION occurs

42
Q

Fluorescent in situ hybridisation ??

A

Take DNA - Denature it
Add fluorescent probe
Allow hybridisation

43
Q

What is the difference between an oncogene and a proto oncogene

A

A proto-oncogene is a normal gene that has many different functions in the cell, if it becomes mutated it will become a oncogene. Think of it like a pre-cursor

Oncogenes are genes which can transform a cell into a tumour cell

44
Q

An extensive genetic test carried out on an unborn baby includes the DNA sequencing on an entire
chromosome. The test reveals a frameshift mutation which leads to a premature stop codon on one
of the alleles of a gene of known function. Explain in general terms what you expect to happen if the
mutant allele is recessive to the wild type allele, and what if the mutant allele was dominant over
the wild type allele

A

Dominant would mean the baby would have the diseased phenotype

Normal would mean the baby had a normal phenotype

45
Q

Give an example of a environmental factor that causes single strand breaks

A

Ionising radiation
X-rays
Anti-tumor drugs
Reactive oxidisation species

46
Q

Give an example of a environmental factor that causes doublestrand breaks

A

Ionising radiation
X-rays
Anti-tumor drugs

47
Q

Give an example of a environmental factor that causes pyrimidine dimers?

A

UV light

Chemicals

MCBG (10 decks)

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