Agarose Gel Electrophoresis Lab Report

DNA lab 2 (temporary): Agarose Gel Electrophoresis

How to pour, load, and run an agarose gel.

MATERIALS

Buffers and Solutions

Agarose solutions (please see Step 3)

DNA staining solution

Electrophoresis buffer

6x Gel-loading buffer Nucleic Acids and Oligonucleotides

DNA samples

DNA size standards

Samples of DNAs of known size are typically generated by restriction enzyme digestion of a plasmid or bacteriophage DNA of known sequence. Alternatively, they are produced by ligating a monomer DNA fragment of known size into a ladder of polymeric forms. METHOD

1. Seal the edges of a clean, dry glass plate (or the open ends of the plastic tray supplied with the electrophoresis apparatus) with tape to form a mold. Set the mold on a horizontal section of the bench.

2.

Prepare sufficient electrophoresis buffer (usually 1x TAE or 0.5x TBE) to fill the electrophoresis tank and to cast the gel.

It is important to use the same batch of electrophoresis buffer in both the electrophoresis tank and the gel.

3. Prepare a solution of agarose in electrophoresis buffer at a concentration appropriate for separating the particular size fragments expected in the DNA sample(s): Add the correct amount of powdered agarose (please see table below) to a measured quantity of electrophoresis buffer in an Erlenmeyer flask or a glass bottle.

Range of Separation in Cells Containing Different Amounts of Standard Low-EEO Agarose

Agarose Concentration in Gel (% [w/v]) Range of Separation