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Flashcards in Molecular Techniques Deck (49)
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1

What produce endonucleases

Bacteria

2

What do restriction enzymes do

Recognise and degrade foreign DNA

3

Which electrode do DNA fragments travel toward in electrophoresis and why

Positive as DNA is negatively charged

4

What sized fragments travel the furthest

Small

5

What does DNA gel electrophoresis do

Separate fragment based on size

6

How are genes cloned

Isolate specific gene with restriction enzymes and insert into plasmid vector to place into bacteria to replicate naturally

7

What are plasmids

Small circular DNA

8

Give an example of when human genes are cloned

To make human insulin

9

What enzyme concerts mRNA into cDNA

Reverse transcriptase

10

What disease is currently being treated using gene therapy

Cystic fibrosis

11

What enzyme is used in PCR and why

Taq Polymerase as it can work at higher temperatures

12

What is the first step in PCR

Heat strands to 95 degrees to denature DNA to give single strands

13

What is the second step of PCR

Cool DNA to 55 degrees to allow DNA primers to bind

14

What is the final step of PCR

Heat to 72 degrees to allows taq Polymerase to bind and add nucleotides from the 3' end.

15

Why is PCR used

Look for mutations, to see if there's a loss or gain of restriction fragments or to see size of product

16

Why can you use PCR to determine if someone has sickle cell anaemia

In sickle cell the mutation changes a restriction site and so this site will not be cut like it would in normal DNA

17

What does PCR result in

Lots of copies of a specific piece of DNA

18

What is protein gel electrophoresis

Separation of proteins

19

What different types of protein electrophoresis are there?

SDS page, isoelectric focussing and 2D page

20

What is SDS page

When proteins are denatured, given a negative charge and separate due to size as they migrate to the positive electrode

21

What is 2D page separation

Separation of proteins based on size and charge

22

What is isoelectric focussing

Separation of proteins based on charge. Protein migrate until they reach a pH equal to their pI

23

What is proteomics

Protein identification

24

How do you carry out proteomics

Digest protein, perform mass spectrometers and identify protein based on peptide sizes

25

What Techniques uses antibodies instead of DNA hybridisation

Western blotting

26

What is ELISA

A technique to detect protein concentration in a mixture by using antibodies to bind to the protein

27

What is an enzyme assay

A technique used to measure enzyme activity by measuring appearance of product or disappearance of substrate

28

The concentration of which enzyme increases during a heart attack

Creatine Kinase

29

What are monoclonal antibodies

Antibodies that can bind to 1 epitope/protein only

30

What are polyclonal antibodies

Antibodies that can bind to multiple epitope in the same antigen