GENETIC ENGINEERING PRACTICAL REPORT

By :
Name : Mellya Rizki Pitriani
Student ID : B1B017031
Entourage : III
Group :4
Assistant : Heri Priyanto

MINISTRY OF RESEARCH, TECHNOLOGY AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
FACULTY OF BIOLOGY
PURWOKERTO
2019
LABORATORY SKILL

A. Aims

The aims of laboratory skill practical class is o equip the students with skills for
using biology molecular laboratory equipment.

B. Materials

The material used in this practical class is worksheet.

The tools used in this practical class are micropippete, microtip, water bath,
vortex, centrifugator, microtube, PCR apparatus, electrophoresis apparatus,
stationary and camera.

C. Methods

The methods used in this practical class are:

1. Tools are prepared
2. Tools that have been prepared are observed
3. Tools that are known are written and their functions on the worksheet.

D. Result and Discussion

Picture 1.1 Microtube Picture 1.2 Micropippete and tip

Picture 1.3 Electrophoresis Apparatus Picture 1.4 Centrifugator

Picture 1.5 PCR Picture 1.6 Vortex

Picture 1.7 UV Transluminator Picture 1.8 Waterbath

 Picture 1.9 PCR for Virus Picture 1.10 Centrifugator
Without Temperature control
Micropipette for taking and homogenizing DNA samples and loading dye. The
use of glass pipettes such as measuring pipettes and goiter pipettes does not have
high accuracy for volumes of less than 1 ml. So that in transferring liquids with small
volumes of less than 1000 microliters, people tend to use micropipets, also
commonly called automatic pipettes. This automatic pipette has better accuracy and
precision than a glass pipette. Each pipette can be set regardless of volume during the
pipette volume range. Micropipettes can be divided into singles, channels,
multichannel and adjustable and fix. There are several types of micropipette brands
on the market such as Gilson, Pipetman, and others (Khopkar, 1990). The key
function of the pipette is to confine the oxygen flux between its two ends, reducing
the lateral spread of the oxygen molecules being detected, while permitting the use of
sensors with larger effective areas positioned further away from source of flux (Gao
et al., 2018).
UV transilluminator for visualization of DNA staining in agarose gels. UV
transilluminators are used to visualize DNA after loading or running in DNA
electrophoresis. Polymerase chain reaction (PCR) is a major laboratory technology in
mocular biology. PCR has a high degree of specificity and sensitivity. With this PCR
technique, in minutes we can produce millions of copies of DNA. A
spectrophotometer is a device used to measure absorbance by passing light with a
specific wavelength on a glass or quartz object called a cuvette. Some of the light
will be absorbed and the rest will be missed. The absorbance value of the light
passed will be proportional to the concentration of the solution in the cuvette
(Martin, 1996).
 PCR, or Polymerase Chain Reaction is used for DNA amplification mostly.
All were used for the molecular biology assay which need multiplication of nucleic
acid in vitro. This become gold standard for quantification of nucleic acid. PCR has
been applied to study genetic cases such as mutation and detecting sensitivity and
resistance of the mutation itself. Gene expression and RNA quantification also been a
subject for studies (Karlin-Neumann & Bizouarn, 2018). DNA sample that has been
amplified then will processed into electrophoresis. Electrophoresis itself used for
examine and separate the sample. The apparatus is consist of comb, tray, gel box or
the chamber, power generator, and electrodes. And the interpretation of DNA were
using UV transluminator (Ramsey et al., 2016).
Water bath is a tool that used for heating materials. Water bath creating a
condition in which the temperature is constant with timer on it. Even the main use of
this tools is for heating, but the purpose may different in every treatment. In some
treatment, lab labor use water bath in proper temperature to aid rehydration.
Centrifuge used for operation involving tissue homogenization to DNA purifying
with the help of enzymes. The result of centrifuge often separating the liquid into 2
parts, natant and supernatant. In some treatment, we only used one centrifuge result,
either natant or supernatant, and the rest of it will be discarded. Centrifuge can be
done either in room temperature or cold temperature (Rogers and Bendich, 1994).
A vortex mixer, or vortexer, is a simple device used commonly in
laboratories to mix small vials of liquid. It consists of an electric motor with the drive
shaft oriented vertically and attached to a cupped rubber piece mounted slightly off-
center. As the motor runs the rubber piece oscillates rapidly in a circular motion.
When a test tube or other appropriate container is pressed into the rubber cup (or
touched to its edge) the motion is transmitted to the liquid inside and a vortex is
created. Most vortex mixers are designed with 2 or 4-plate formats, have variable
speed settings ranging from 100 to 3,200 rpm, and can be set to run continuously, or
to run only when downward pressure is applied to the rubber piece (Martin, 1996).

REFFERENCE

Gao, Y., Li. B., Riju, S., Adam, F., Ben, P., Zulfiya, O., Yury, G., Gary, F., 2018.
Perfusion Double-Channel Micripipette Probes for Oxygen Flux Mapping with
Single-cell Resolution. Beilstein Journal of Nanotechnology, Vol 9, pp. 850-
860.
Karlin-Neumann, G., & Bizouarn, F., 2018. Entering the Pantheon of 21st Century
Molecular Biology Tools: A Perspective on Digital PCR. Digital PCR, 3–10.

Khopkar, S. M., 1990. Konsep Dasar Kimia Analitik. Jakarta: UI Press.

Martin, 1996. Gel Electrophoresis: Nucleic Acid. Oxford: Bios Scientific Publisher.
Rogers, S. O., & Bendich, A. J., 1994. Extraction of total cellular DNA from plants,
algae and fungi. Plant Molecular Biology Manual, 183–190.

Ramsey, K. A., Rushton, Z. L., & Ehre, C., 2016. Mucin Agarose Gel
Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins.
Journal of Visualized Experiments, (112), pp. 1-6.