Introduction to Molecular Techniques Flashcards
Name 3 ways in which DNA can be analysed at a gene level
- Restriction enzymes
- DNA gel electrophoresis
- Polymerase chain reaction (PCR)
What are restriction enzymes?
Molecular scissors that each recognise a specific DNA sequence
What class of enzymes do restriction enzymes belong to?
Endonucleases
What characteristic do most restriction enzyme DNA sequences have?
Palindromes
What 4 components are required for DNA gel electrophoresis?
- Gel
- Buffer
- Power supply
- Stain
How does DNA gel electrophoresis separate DNA?
- DNA is negatively charged and will move towards the anode
- DNA fragments are separated by size
What carriers are used for gene cloning and what does this technique allow genes to do?
- Uses bacterial plasmids as vectors
- Carry genes which can replicate individually
Describe the 4 steps involved in the process of gene cloning
- Isolate the relevant gene after digestion with restriction enzymes
- Insert gene into vector using DNA ligase
- Introduce recombinant DNA into a host cell
- Identify and isolate the clone
Name 3 uses of gene cloning as a DNA analysis technique
- Can make useful proteins (insulin)
- Find our what genes do
- Genetic screening
- Gene therapy
What is Polymerase Chain Reaction (PCR)?
Allows amplification of target DNA sequences using thermostable DNA polymerase
Which DNA polymerase is used in Polymerase chain reaction?
Taq
Name 3 components of the polymerase chain reaction technique
- Forward and reverse primer
- Thermostable DNA polymerase
- Temperature cycles
Name 2 uses of polymerase chain reaction DNA analysis
- Amplify a specific fragment
- Investigate single base mutations
- Investigate small deletions or insertions
- Investigate variation and genetic relationships
Which mechanism is used in protein gel electrophoresis?
Uses the isoelectric point of proteins to separate them by movement towards an anode or cathode
How are proteins separated by protein gel electrophoresis?
Size, shape or charge
