Lecture 15 Flashcards
How does DNA sequencing work?
- DNA is denatured and a template is used
- A primer is the annealed to the 5’ end of the DNA
- Primed DNA is dispersed to 4 vessels
- DNA pol is added
- 4 dNTP to each
- One type of ddNTP is then added to each
- pol attached the dNTP to the template
- Once, ddNTP is paired, the sequence is terminated because it lacks hydroxyl group at the 3’
- different fragments form
- pol gel is used
- DNA migrates from - to +
Why is polyacrylamide gel used rather than agarose?
Because it has high resolving power and can separate DNA strands that differ in length by 1 base pair
Why are long reads preferred for cancer research?
Allows for seeing changes that could not be seen in old tech. Shows progression and is cheaper
What are transposons?
Mobile genetic elements or “jumping genes” from one location of the DNA to the next
Who discovered transposons?
Barbara McClintock
What are the two classes of transposons?
Class 1 & 2
What is class 1?
Move from copy RNA transcription and then copied back to DNA from reverse transciption, transcriptase. DNA is then inserted back to genome
What is class 2?
No RNA intermediate. Remains in DNA form. Encode tranposase.
What does transposase do?
It will cut transposon and insert it somewhere else in the DNA. Uses cut and paste mechanism
