Comb/ well forming templates

 Comb is placed at one end of the Gel casting tray before casting the gel to make wells
(sample slots) for the sample to be pipetted into.
 The dimension of the tooth of the comb defines the dimensions of the well. The more the
number of tooth in a comb, the smaller is the capacity of the wells.
 Once the gel has cooled and solidified (it becomes opaque rather than clear) the comb is
removed.

Gel sealing tape

 Common type of lab tape such as Time tape or VWR tape.

 It is used for sealing the edges of clean, dry glass plate or the plastic tray of the
electrophoresis apparatus to form a mould in which the molten gel is poured for the casting
of the gel.

Power pack

 It is used to supply power of upto 500 V and 200mA

 The power pack has two lead, a black lead, the negative lead and a red lead, the positive
lead. The negative electrode is connected at the end near the well and the positive lead is
connected to the other end. After loading the sample mixed with dye, the leads are
connected and a voltage of 1-5 V cm-1 is applied across the terminals.
 When the leads are connected properly and voltage is applied, bubbles start erupting at the
base of the anode and cathode.
 When the power is applied, the DNA astarts migrating towards the anode while EtBr
migrates towards the cathode.

Gel loading buffer (follow the material provided)

Gel loading buffers are mixed with the samples before loading into the slots of the gel. The
gel loading buffers serve three purposes:

i. They increase the density of the sample, ensuring that the DNA sinks evenly into the well
ii. They add colour to the sample, thereby simplifying the loading process
iii. They contain dye that migrates towards the anode in an electric field.
 Micropipette

 Micropipettes are used to transfer small volumes of DNA samples, usually down to 0.1 uL
into the wells of the casted gel.
 Micropipette is also used to mix the dye, Ethidium dibromide with the DNA samples by
constantly pressing and releasing the pipette, while mixing though the pipette is not
released completely but is pressed and released synchronously.

Micropipettes differ in size and volume dispensed. Micropipettes use a disposable pipette
tip to aspirate liquid. A new tip is utilized for every sample in order to prevent cross
contamination.

Molecular weight size markers (DNA ladder):

 DNA ladder is a set of size standards ( generated by restriction enzyme digestion of a

plasmid or bacteriophage DNA of known sequence) that are used to identify the
approximate size of a molecule run on a gel during electrophoresis, using the principle that
molecular weight is inversely proportional to migration rate through a gel matrix.
 A stock solution of the DNA ladder is prepared by dilution with a gel loading buffer.

Electrophoretic chamber

Electrophoresis chamber is a device used to generate a potential difference ( V) across two

ends of a gel in an electrolyte/buffer solution. This potential difference causes the
movement of charged molecules (such as negatively charged nucleic acids) towards the
anode, positive electrode through the gel which results in the separation of the DNA
molecules by size.