Sequencing Technology Overview

Question 1

How does Sanger sequencing technology work?

Answer 1

DNA polymerase attaches to the primer an adds nucleotides as it walks along the primer
ome of the nucleotides that add cannot be extended upon. As you expand, there are fewer and fewer templates, but we have information on what the last based added was. So short fragments can be ran out on a gel n based on the color of the nucleotide, you can read the sequence of the DNA based on where it lands on the gel

Question 2

What is single end sequencing?

Answer 2

just read on end of the DNA at a time , so if you have a 150bp genomic fragment, you do 100 bp single sequencing and read for first 100 bp. So the information you get back is just the first 100bp of that fragment.

Question 3

What is double end sequencing?

Answer 3

A neat trick you can do is read both ends of the fragment at the same time. Ex: If you have a 800bp fragment, read the first 100bp in the forward direction and then you can go and read the other 100bp in the reverse direction. The data that you get back has the first and last 100 bp of the DNA. You know there’s 600bp gap in between , but you know what each side of the DNA looks like. This is helpful when you’re aligning stuff to the genome or when you’re looking for things like duplications , deletions, etc.