REAGENTS OF GEL ELECTOPHORESIS 2021

ALIYA NAWAZ

GENETICS LAB ASSIGNMENT

REGISTRATION NO: SP20-BSI-006 | INSTRUCTOR: DR

MALEEHA AZAM
 GENETICS LAB ASSIGNMENT

GEL ELECTROPHORESIS REAGENTS

BACKGROUND:
Gel electrophoresis is the most laboratory technique used to separate biological
molecules based on their size. This process is helpful in a range of research
applications such as identifying unknown samples plus mainly used for the
separation of DNA molecules.

REAGENTS:
The reagents used in practical of Gel Electrophoresis are as under:
• Agarose Powder • Buffer Stocks • Distilled Water • Loading dye • DNA Ladders
• DNA Stains

AGAROSE:
 Agarose is purified form of agar. It is made up of polymer that is comprised
of repeating molecules of 2 forms of galactopyranose.
 It is ideal for gel electrophoresis because agar has no net charge and
therefore, it will not be able react with running buffers and molecules can
never be utilized in gel running process.

BUFFER STOCK:
TBE:
TBE stands for tris-burate EDTA that is a running buffer used for separating small
fragments of DNA. It gives higher resolution of small DNA fragments. It has
higher buffering capacity and lower conductivity. It is used for running gel at high
voltage.
 EDTA: is chemical which chelates to heavy metals like Zn++ and to
divalent cations that is Mg++. It is added at low concentration to inactivate
any trace of heavy metal that will inhibit the reaction. The restriction
enzyme buffer is used as for digesting DNA
Tris-HCl:
 It is a buffer which is commonly used to maintain pH which range over 7 to
9 during addition of acid and base.

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 GENETICS LAB ASSIGNMENT

 This is because DNA is very sensitive to pH change and pH change may

cause damage to DNA. Therefore, tris-HCl is used.

DISTILLED WATER:
 It is used in all biological molecules solution because tap water consist of
heavy metals, traces of salts, chlorine, fluorine and dirt that can affect
biological molecules under study.

ETHIDIUM BROMIDE (DNA STAIN):

 Ethidium is chemical commonly used in gel electrophoresis as non-
radioactive marker for identifying and visualizing DNA band in
electrophoresis.
 It fluorescence readily having reddish brown color during exposing to UV
light, intensifying almost 20 folds after binding to DNA.
 The traces of heavy metals present in tap water can damage DNA and can
inhibit some enzymes.

LOADING DYE:
The loading dye is a mixture of glycerol and three other dyes that move through
the gel at differing rates. Loading dyes impart colour to the samples, which helps
in visualizing the loading process. It also increases the density of the sample.

NaCl:
 Sodium chloride salt help in adjusting the ionic strength of solution that
should be equivalent to that inside the cell and they interact with charged
groups on surface of DNA.