DNA sequencing technology CMSC 828N, Fall 2008

Restriction enzymes

Library construction First, do size selection run the DNA on a gel Cut out a swath of DNA for the desired size repeat [Switch to cloning slides here]

After cloning Lyse (cut open and kill) E. coli with plasmids Amplify sequences directly from built-in primers

DNA sequencing

Dideoxy terminators (modified DNA)

Extend the sequences

Run sequences through a gel

Fluorescent dyes

Animated gel separation

Multiple “lanes” at one time

Chromatogram data

PCR Amplify any DNA using 2 primers Details here

PCR animation