Home Blast Tutorial Part 1: DNA Sequencing: A. Viewing a Sequence

BLAST Tutorial

Part I:
DNA Sequencing:
A. Viewing a Sequence

B. Reading a DNA Sequence
C. "Calling" Your Own Sequence

An Ara-mini Tn-10 translational fusion was analysed by PCR, using the Ara/LacZ primers that you used on RDM Day 3. A band was seen with the AraC/LacZ primer set. The AraC/LacZ PCR product was sequenced in order to confirm that the mutant was indeed an AraC translational fusion, and to identify precisely where in ara C the transposon had inserted.

The image on the right is the raw output from a sequencing gel, using the "dye terminator method." Eight lanes are shown, representing four reactions carried out in duplicate (duplicate lanes are right next to each other, so they may appear as one large lane).

The lanes are loaded as follows:

Lanes 1-4 are the same as lanes 5-8, except that they use only 33% of the recommended sequencing kit reagents. "AraC" indicates that the AraC primer was used to sequence, whereas "LacZ" samples used the LacZ primer for sequencing.