Module 6: Manipulating genomes Flashcards
What are exons
Coding DNA
What are introns
Non coding DNA
What is sataltide DNA
short DNA sequenes that are repeated
What is a mini satalite
DNA sequence that are 20-50 pairs long
repeated 50-100 times
What are mircosatalites
DNA sequence 2-4 bases
repeated 5-15 times
Where do we find satalites
on the same area of a chromosome
What is the principle of DNA profiling
different people have different number of repeats as they have differnet satalite patterns
What do we use DNA profiling for
Paternal testing
Looking for diseases
Forensics
What are the stages of DNA profiling
- Extract DNA
- Amplify DNA though PCR
- Add restriction endonucleuases
- Gel electrophresis
- Southern Blotting
- Hybridisation
What goes into a thermal cycler (PCR machine)
Primers
Nucleotides
Taq polymerase
What happens during PCR
- temp is at 95 degrees, the hydorgen bonds between base pairs break
- temp lowered to 55, primers anneal at the end of the DNA strand
- Temp increased to 72, free DNA nucelotides bind onto exposed base pairs and taq polyermase reforms the sugar phosphate backbone
Why do we use Taq polymerase
It is found in hot springs
it can survive at high temperatures and it won’t denature
therefore it can be used in many cycles
What is a primer
short piece of single stranded DNA that is complimentary to the bases at the start of the DNA fragement you want